| Research (if you don't see a menu on the left, use this
link)
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The
main objective of the
Department (CBE
= Cristalografía
y Biología
Estructural
= Crystallography and Structural Biology) is to
interpret biological phenomena in terms of structural studies at the
atomic,
molecular and supramolecular level, trying to
understand the
basic processes of life and to use this knowledge to solve
biotechnological and
biomedical problems. To achieve this objective, we combine and develop molecular biology techniques, high throughput crystallization technologies, crystallographic techniques and data-base mining. The research of the Department is focused in several interrelated lines, including structural enzymology, structure and disease, protein-lipid interaction and structure of complex modular systems. To see some short movies about the CBE department, our labs and some of our projects, follow this link. |
| Projects | Main
current projects:
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The
Department is a partner of the Spanish Project
called "La
Factoria", a collaborative project of nine Spanish
research groups to create an integrated platform for research and
services in crystallization and crystallography. |
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Structural Biology of Cellular Stress Response Project Leader: Armando Albert |
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Arabidopsis thaliana OST1 belongs to the SnRK2 family of protein kinases. This family and their interacting regulators, the protein phosphatases type 2C (PP2C) function together in decoding plant cell signals elicited by different environmental stimuli. Available data suggest a general mechanism in which the interaction of SnRK2 and PP2C regulates the phosphorylation state and activity of SnRK2 and this in turn, regulates the phosphorylation state and the activity of various ion transporters and transcription factors. OST1 is activated by osmotic stress as well as by abscisic acid (ABA). The recent identification of the PYR family of ABA receptors as inhibitors of the PP2C phosphatases has provided the basis of the ABA dependent regulation of this family of kinases. At molecular level, structural studies have shown that sensing ABA involves a molecular rearrangement of the PYR receptor that provides a site for interaction with PP2Cs active site. This information helped to explain how the pathway captures the ABA message but it did not provide insights about how SnRK2 handles the signal. The structure of OST1 provides the basis for the ABA dependent and ABA independent regulation of the kinase activity and a mechanism for the control of the opposing phosphatase-binding and kinase activities. Click here, or on the image, to see an animated gif. Selected references: Yunta, C; Martínez-Ripoll, M; Jian-Kang Zhu, J-K; Albert, A Journal of Molecular Biology (2011) 414, 135–144 (doi:10.1016/j.jmb.2011.09.041) Yunta, C; Martínez-Ripoll, M; Albert, A Acta Crystallographica (2011) F67, 364–368 (doi:10.1107/S1744309110053807) |
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The crystal structure of the binary complex of Ca-SOS3 with the C-terminal regulatory moiety of SOS2 resolves central questions regarding the dual function of SOS2 as a kinase and a phosphatase-binding protein. A comparison with the structure of unbound SOS3 reveals the basis of the molecular function of this family of kinases and their interacting calcium sensors. Furthermore, our study suggests that the structure of the phosphataseinteraction domain of SOS2 defines a scaffold module conserved from yeast to human. Selected references: Sánchez-Barrena, MJ; Fujii, H; Angulo, I; Martínez-Ripoll, M; Zhu, J-K; Albert, A Molecular Cell (2007) 26, 427-435 (doi:10.1016/j.molcel.2007.04.013) |
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X-ray
crystallography shows that SOS3 displays a calmodulin-like fold but
forms dimers. Analytical ultracentrifugation and circular dichroism
studies confirm that calcium not only induces a conformational change
(as in the case of calmodulin) but also promotes dimerization of SOS3.
It is proposed that dimerization of SOS3 (mediated by the C-terminal
part) duplicates the efficiency of membrane binding (mediated by
myristoylation and a polylysine stretch at the N-terminus). Membrane
binding of SOS3 is essential for its biological function, probably by
recruiting protein kinase SOS2 to its transporter substrates such as
the H+/Na+ antiporter
SOS1.
http://www.facultyof1000.com/;
Selected by Ramon Serrano, Evaluated 25 Feb 2005) Selected references: Sánchez-Barrena, MJ; Martinez-Ripoll, M; Zhu,J-K; Albert, A Journal of Molecular Biology (2005), 345, 1253-1264 [PDF-copy] |
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Enzymes such as yeast HAL2 and plant homologs, which are proteins belonging to the inositol monophosphatase family and are sodium inhibited. It is known that HAL2 is the first cellular target of sodium under salt stress conditions. Selected references: Patel, S; Martinez-Ripoll, M; Blundell, TL; Albert, A Journal of Molecular Biology (2002), 320, 1087-1094 [PDF-copy] Albert, A; Yenush, L; Gil-Mascarell, MR; Rodriguez, PL; Patel, S; Martinez-Ripoll, M; Blundell, TL; Serrano, R Journal of Molecular Biology (2000), 295, 927-938 [PDF-copy] |
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Domains from enzymes or peptidic regulators of the cellular pathways involved in the response to the salt stress. HAL3 is a plant flavoprotein that regulates a novel signaling pathway involved in cell growth and stress response. Selected references: Albert, A; Martinez-Ripoll, M; Espinosa-Ruiz, A; Culiañez-Macia, F; Serrano, R Structure with Folding and Design (2000) 8, 961-969 [PDF-copy] [Further information on this subject in a pdf file] |
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Inositide Signalling Structural Biology Project Leader: Beatriz González Pérez |
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| The research is focused on the Structural
Biology of the Inositide Signalling. The inositides are second
messengers that can be found in the cell citosol, (Inositol Phosphates
or IPs), or tethered to the cell membrane (PIs, Phosphatidyl Inositides
or lipid inositides).
These second messengers regulate many key events, having crucial roles
in Ca2+ liberation from internal stores, ionic
channels regulation, vesicle trafficking, endocytosis, DNA reparation,
telomere length, etc. We are looking at the kinases in charge of
regulating the inositide compounds levels, with the purpose of
understanding their substrates specificity, catalytic and regulatory
mechanism, protein-protein interactions... As a final goal, we are
interested in drug design to be able to find appropriate therapies to
treat diseases originated by their wrong regulation. |
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Phytate or InsP6 is a key compound in cell and plant biology. InsP6 is involved in essential cellular processes as RNA export or DNA editing. In plants, InsP6 accumulates in seeds, beans and tubers. This makes grain-based diets harmful for human health, since an excess of InsP6, a potent chelator of essential ions as Zn2+ and Fe2+, lead to malnutrition and diseases as anaemia. In addition, many animals are unable to digest the phytate phosphorus present in their feed, which is transferred as manure that leads to pollution of waterways. For these reasons, the animal feedstuffs industry adds an enzyme called phytase to the feedstuff, which allows animals to absorb the phosphorus from phytate. This is a costly process, and so the industry needs to identify low-phytate varieties of crops such as maize, rice, wheat, barley and soya bean. Our work shows the molecular basis of how phytate is synthesised in cells by InsP5 2-K. The results reveal how the enzyme recognises its substrates through a novel structural region (CIP-lobe), and the elements involved in the myo-inositol isomer selection. All these findings represent an important tool to design inhibitors for the enzyme, what have potential applications in biomedicine and animal feedstaff industry, for example in designing crops with low phytate levels. Selected references: Inositol 1,3,4,5,6-pentakisphosphate 2-kinase is a distant IPK member with a singular inositide binding site for axial 2-OH recognition González, B; Baños-Sanz, J; Villate, M; Brearley, C; Sanz-Aparicio, J Proceedings of the National Academy of Sciences (2010) 107, 9608-9613 (doi:10.1073/pnas.0912979107) Crystallization and preliminary X-ray diffraction analysis of inositol 1,3,4,5,6-pentakisphosphate kinase from Arabidopsis thaliana Baños-Sanz, J.I., Villate, M., Sanz-Aparicio, J., Brearley, C.A. and González B. Acta Crystallographica (2010) F66, 102-106 (doi:10.1107/S1744309109051057) |
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Phosphatidyl Inositol Kinases (PI3Ks) are a family of enzymes involved in cancer and other important diseases, therefore being a good target for drug design. The structure of PI3K in complex with different inhibitors allowed us to determine structure regions that control the selectivity and potency of the inhibition. Our last work looks for inhibitors with dual selectivity against two proteins among the most pursued targets in cancer, PI3K and tyrosine kinases. The aim is to overcome the problems of the resistance generated against the tyrosin kinases drugs used in cancer therapy. The structure of the complexes between the enzymes and these inhibitors reveals that this dual selectivity is controlled by a hydrophobic pocket conserved in both enzyme classes. The results yield a possible future cancer drug and the structural basis to continue the rational drug design for these targets. Selected references: Apsel, B; Blair. JA; González, B; Nazif, TM; Feldman, ME; Aizenstein, B; Hoffman, R; Williams, RL; Shokat, KM, Knight, ZA Nature Chemical Biology (2008) 4, 691-699 Knight, ZA; González, B; Feldman ME; Zunder, ER; Goldenberg, DD; Williams, O; Loewith, R; Stokoe, D; Balla, A; Toth, B;, Balla, T; Weiss, WA; Williams, R;. Shokat, KM Cell (2006) 125, 733-747 [PDF-copy] Teo, H; Perisic, O; González, B; Williams, RL Developmental Cell (2004) 7, 559-569 [PDF-copy] |
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We have solved
the structure of a first inositol kinase acting on a soluble inositol
(the IP3 3-kinase). This enzyme uses IP3
as a substrate, a second messenger involved in Ca2+
release from internal stores, and has been shown to be essential in
T-lymphocyte development. The structure shows that this family of
enzymes conserves fold elements with the protein kinase family, binding
ATP in a similar fashion. Nevertheless, the inositide substrate is
bound in a new lobe formed by four α-helices. Thoroughly
alignments among all the IP3 3-K family members
lead us to predict that
the inositide binding lobe is unique in IP3 3-K,
presenting the other
members only one α-helix in this lobe. This is coherent with
the
necessity of a more constrained active site for IP3
3-K due to its
greater specificity. Selected references: González, B; Schell, MJ; Letcher, AJ; Veprintsev, DB; Irvine, R; Williams, RL Molecular Cell (2004) 15, 689-701 |
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Structural Biology of Bacterial Pathogenesis Project Leader: Juan A. Hermoso |
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| Streptococcus pneumoniae is a human
pathogen responsible for some of the infection diseases with higher
index of mortality and morbidity worldwide. Our work aims the study of
the structural biology of host-pathogen interactions in infections by
Streptococcus pneumoniae. The objective is to provide the basic
knowledge needed for future development of novel prevention, diagnostic
and treatment tools against pneumococcal infections. Experimental
approaches include the use of protein engineering and X-ray diffraction
techniques to the study of virulence mechanisms mediated by the
pneumococcal surface proteins such as the so called choline-binding
proteins (CBP). We also focus on the host proteins involved in pathogen
recognition and on the structural characterization of novel endolysins
(phage-encoded enzymes that break down bacterial peptidoglycan) with
potential application as antibacterial agents (enzybiotics). |
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AmpD is a cytoplasmic peptidoglycan amidase involved in bacterial cell wall recycling and in the induction process of class C β-lactamases. Crystal structures of AmpD revealed, for the first time, the presence of an open/active and a closed/inactive conformation. The activation mechanism involves large structural rearrangements (17 Å movement) in one third of the entire protein. This activation mechanism might be representative of a regulatory process for other intracellular members of the bacterial amidase_2 family of enzymes. Click on the image (or on this link) to see a movie showing the whole activation mechanism. Selected references: Carrasco-López, C; Rojas-Altuve, A; Zhang, W; Hesek, D; Lee; M; Barbe, S; André, I; Ferrer, P; Silva-Martin, N; R. Castro, G; Martinez-Ripoll, M; Mobashery, S; Hermoso, JA Journal of Biological Chemistry (2011) 286, 31714-31722 (doi:10.1074/jbc.M111.264366) |
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The crystal structure of the first endolytic peptidoglycan lytic transglycosylase MltE from Escherichia coli is reported herein. The degradative activity of this enzyme initiates the process of cell wall recycling, which is an integral event in the bacterial existence. The structure sheds light on how MltE recognizes its substrate, the cell wall peptidoglycan. It also explains the ability of this endolytic enzyme to cleave in the middle of the peptidoglycan chains. Furthermore, the structure reveals how the enzyme is sequestered on the inner leaflet of the outer membrane. Selected references: Artola-Recolons, C; Carrasco-López, C; Llarrull, L; Kumarasiri, M; Lastochkin, E; Martínez de Ilarduya, I; Meindl, K; Usón, I; Mobashery, S; Hermoso, JA Biochemistry (2011) 50, 2384-2386 (doi:10.1021/bi200085y) Artola-Recolons, C; Llarrull, L; Lastochkin, E; Mobashery, S; Hermoso, JA Acta Crystallographica (2011) F67, 161-163 (doi:10.1107/S1744309110049171) |
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The first structure of a pneumococcal autolysin, that of LytC, has been solved in ternary complex with choline and a pneumococcal peptidoglycan fragment. The active site of the hydrolase module is not fully exposed but oriented towards the choline-binding module accounting for its unique in vivo features in peptidoglycan hydrolysis, its activation and its regulatory mechanisms. Due to the unusual hook-shaped conformation of the multimodular protein, it is only able to hydrolyze non-crosslinked peptidoglycan chains, an assertion validated by additional experiments. These results explain the activation of LytC by CbpD in fratricide, a competence-programmed mechanism of predation of noncompetent sister cells. The results provide the first structural insights into the critical and central function that LytC plays in pneumococcal virulence and explain a long-standing puzzle of how murein hydrolases can be controlled to avoid self-lysis during bacterial growth and division. Enjoy a short movie through this link, explaining how CbpD activates LytC in fratricide... Following this other link you can also see a longer video, illustrating two aspects on the pneumococcal virulence, as based on two articles concerning the structure and function of some pneumococcal enzymes. Selected references: Pérez-Dorado, I; González, A; Morales, M; Sanles, R; Striker, W; Vollmer, W; Mobashery, S; García, JL; Martínez-Ripoll, M; García, P; Hermoso, JA Nature Structural & Molecular Biology (2010) 17, 576-582 (doi:10.1038/nsmb.1817) Pérez-Dorado, I; Sanles, R; González, A; García, P; García, JL; Martínez-Ripoll, M; Hermoso, JA Acta Crystallographica (2010) F66, 448-451 (doi:10.1107/S1744309110006081) |
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Pneumococcal surface proteins are the major pool of virulence factors and represent key players in pneumococcal adhesion, colonisation and virulence. Both the three-dimensional structure and the function of choline-binding protein F (CbpF), one of the most abundant proteins in the pneumococcal cell wall, are now reported. While its C-terminal module is involved in cell wall binding, the N-terminal module is critical for inhibition of the autolytic LytC muramidase, providing a regulatory function for pneumococcal autolysis with relevant implications in pivotal processes such as competence and allolysis. Selected references: Molina, R; González, A; Stelter, M; Pérez-Dorado, I; Kahn, R; Morales, M; Campuzano, S; Campillo, NE; Mobashery, S; García, JL; García P; Hermoso, JA EMBO Reports (2009) 10, 246-251 (doi:10.1038/embor.2008.245) Molina, R; González, A; Moscoso, M; García, P; Stelter, M; Kahn, R; Hermoso, JA Acta Crystallographica (2007) F63, 742-745 (doi:10.1107/S1744309107035865) |
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The
crystal structure of the pneumococcal phosphorylcholine esterase, Pce
(602 aminoacids), revelaed Pce could selectively modify the
distribution of the phosphoryl choline moieties on the bacterial
surface. Thus Pce should impair the ability of human defense system to
efficiently bind the bacteria, and would provide a mechanism for
pneumococci escaping the immune attack. Besides, we have verified the ability of Pce to hydrolyze PAF (human platelet-activating factor) a pivotal first messenger of the inflammatory processes suggesting that this enzyme has other functions during infection. This finding opens a new scenario on the role that Pce may play in the mechanism of pneumococcal adherence and invasiveness. Enjoy a short movie through this link, explaining the role of Pce in pneumococcal camouflage... Following this other link you can also see a longer video, illustrating two aspects on the pneumococcal virulence, as based on two articles concerning the structure and function of some pneumococcal enzymes. Selected references: Hermoso, J; Lagartera, L; Gonzalez, A; Stelter, M; Garcia, P; Martinez-Ripoll, M; Garcia, JL; Menendez, M Nature Structural and Molecular Biology (2005) 12, 533-538 [PDF-copy] Lagartera, L; González, A; Hermoso, JA; Saíz, JL, García, P; García, JL; Menéndez, M Protein Science (2005), 14, 3013-3024 Lagartera, L; Gonzalez, A; Stelter, M; Garcia, P; Garcia, JL; Menendez, M; Kahn, R; Hermoso, JA Acta Crystallographica (2005) F61, 221-224 |
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The
bactericidal activity of bacteriophages has been used to treat human
infections for years as an alternative or a complement to antibiotic
therapy. Nowadays, endolysins (phage-encoded enzymes that break down
bacterial peptidoglycan at the terminal stage of the phage reproduction
cycle) have been used successfully to control antibiotic-resistant
pathogenic bacteria in animal models (enzybiotics). Their cell wall
binding domains target the enzymes to their substrate, and their
corresponding catalytic domains are able to cleave bonds in the
peptidoglycan network. The first insight on this structural knowledge
has been very recently provided by the X-ray crystal structures of the
Cpl-1 lysozyme in complex with three bacterial cell wall PG analogues.
These findings provide the first structural evidence on recognition of
the peptidoglycan and shed light on the discrete events of cell wall
degradation by Cpl-1. Selected references: Hermoso, JA; García, JL; García, P Current Opinion in Microbiology (2007) 10(5), 461-472 Pérez-Dorado, I; Campillo, NE; Monterroso, B; Hesek, D; Lee, M; Páez, JA; García, P; Martínez-Ripoll, M; García, JL; Mobashery, S; Menéndez, M; Hermoso, JA Journal of Biological Chemistry (2007) 282, 24990-24999 Buey, RM; Monterroso, B; Menéndez, M; Diakun, G; Chacón, P; Hermoso, JA; Díaz, JF J. Mol. Biol. (2007) 365, 411-424 Hermoso, JA; Monterroso, B; Albert, A; Galán, B; Ahrazem, O; García, P; Martínez-Ripoll, M; García, JL; Menéndez, M. Structure (2003), 11, 1239-1249 [PDF-copy] Monterroso, B; Albert, A; Martínez-Ripoll, M; García, P; García, JL; Menéndez, M; Hermoso, JA Acta Crystallographica (2002) D58, 1487-1489 [PDF-copy] [Further Information] |
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Structural Biology of Carbohydrate Active Enzymes Project Leader: Julia Sanz-Aparicio |
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| The carbohydrate active enzymes catalyse the synthesis and breakdown of glycosidic bonds in carbohydrates that are in the form of glycoproteins, glycolipids and polysaccharides. This wide group of enzymes accounts for 1-3 % of the proteins encoded by genomes of most organisms, and are essential in a wide variety of biological processes ranging from energy storage and utilization to highly specific signalling roles. Structural studies of these enzymes and their complexes with substrate analogues and inhibitors reveal structural details that correlate with mechanism and give valuable information that helps us to understand their function at the molecular level. | |
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Our work has focused on bacterial glycosidases involved in the degradation of small olligosaccharides and also in the cleavage of the more complex plant cell wal polysacharide xylan. Some of these enzymes are modular comprising catalytic modules appended to one or more accesory domains that promote substrate binding. Our aim is to have a deep insight into the machinery involved in specificity and the improvement of their stability has also been pursued by structure-based protein engineering. Recent work has started on yeast β-fructofuranosidases (invertases), involved in the preparation of prebiotic oligosacharides (functional foods), and β-galactosidases (lactases) and α-galactosidases from different eukaryote organisms. The structural analysis of these enzymes provides the molecular basis to improve their properties such as glycosylating activity, stability and catalytic efficiency, which is most interesting for biotechnological purposes. Furthermore, it gives a deeper insight into the structural features that rule modularity, a pivotal property within glycosidases crucial to many biologically relevant processes. Selected references: Pereira-Rodríguez, A; Fernández-Leiro, R; González-Siso, MI; Esperanza Cerdán, M; Becerra, M; Sanz-Aparicio, J Journal of Structural Biology (2012) 177, 392–401 (doi:10.1016/j.jsb.2011.11.031) Lafraya, A; Sanz-Aparicio, J, Polaina, J; Marín-Navarro, J Applied Environmental Microbiology (2011) 77, 6148-6157 (doi:10.1128/AEM.05032-11) Alvaro-Benito, M; Abreu, M; Portillo, F; Sanz-Aparicio, J; Fernández-Lobato, M Applied Environmental Microbiology (2010) 76, 7491-7499 (doi:10.1128/AEM.01614-10) Polo, A; Linde, D; Estévez, M; Fernández-Lobato, M; Sanz-Aparicio, J Acta Crystallographica (2010) F66, 1441-1444 (doi:10.1107/S1744309110029192) Fernández-Leiro, R; Pereira-Rodríguez, A; Esperanza Cerdán, M; Becerra, M; Sanz-Aparicio, J Journal of Biological Chemistry (2010) 285, 28020-28033 (doi:10.1074/jbc.M110.144584) Alvaro-Benito, M; Polo, A; González, B; Fernández-Lobato, M; Sanz-Aparicio, J Journal of Biological Chemistry (2010) 285, 13930-13941 (doi:10.1074/jbc.M109.095430) Gallardo, O; Pastor, FJ; Polaina, J; Díaz, P;Vogel, P; Isorna, P; González-Pérez, B; Sanz-Aparicio, J Journal of Biological Chemistry (2010) 22, 2721-2733 (doi:10.1074/jbc.M109.064394) Pereira-Rodríguez, A; Fernández-Leiro, R; González-Siso, M; Cerdán, E; Becerra, M; Sanz-Aparicio, J Acta Crystallographica (2010) F66, 297-300 (doi:10.1107/S1744309109054931) Fernández-Leiro, F; Pereira, A; Cedán, E; Becerra-Fernández, M; Sanz-Aparicio, J Acta Crystallographica (2010) F66, 44-47 (doi:10.1107/S1744309109047794) Polo-Rivas, A; Alvaro-Benito, A; Fernández-Lobato, M; Sanz-Aparicio, J Acta Crystallographica (2009) F65, 1162-1165 (doi:10.1107/S174430910903938) Pastor, FJ; Gallardo, O; Sanz-Aparicio, J; Díaz, P in Industrial Enzymes: Structure, Function and Applications (2007), Chpt. 5, 65-82, Springer NL, Polaina, J; MacCabe, Andrew P (Eds.), XII, 642 p., Hardcover. ISBN: 978-1-4020-5376-4 León, M; Isorna, P; Menéndez, M; Sanz-Aparicio J; Polaina, J Protein Journal (2007) 26, 435-444 Isorna, P; Polaina, J; Latorre-García. L; Cañada, FJ; González, B; Sanz-Aparicio, J Journal of Molecular Biology (2007) 371, 1204-1218 González-Blasco,G; Sanz-Aparicio,J; González-Pérez,B; Hermoso,JA; Polaina,J Journal of Biological Chemistry (2000), 275, 13708-13712 Sanz-Aparicio, J; Hermoso, J; Martínez-Ripoll, M; Lequerica, JL; Polaina, J Journal of Molecular Biology (1998), 275, 491-502 [PDF-copy] Sanz-Aparicio, J; Hermoso, JA; Martínez-Ripoll, M; González-Pérez, B; Lopez-Camacho, C; Polaina, J Proteins: Structure, Function and Genetics (1998), 33, 567-576 Sanz-Aparicio, J; Romero, A; Martinez-Ripoll, M; Madarro, M; Flors, A; Polaina, J Journal of Molecular Biology (1994), 240, 267-270 [PDF-copy] |
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Enzymes from Lactic Acid Bacteria Project Leader: Jose M. Mancheño |
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Catalytic mechanism of p-coumaric acid
decarboxylase from Lactobacillus plantarum The p-coumaric acid decarboxylase from the lactic acid bacterium Lactobacillus plantarum has an internal amphipathic cavity in which the active site is located. Combining crystallographic, molecular biology, and biochemical studies a novel catalytic mechanism of decarboxylation has been proposed together with revealing the existence of conformational changes associated with the catalysis. These results envisage new biotechnological applications for this family of enzymes. Selected references: Rodríguez, H; Angulo, I; de las Rivas, B; Campillo, N; Páez, JA; Muñoz, R; Mancheño, JM Proteins: Structure, Function, and Bioinformatics (2009) 78, 1662-1676 (doi:10.1002/prot.22684) Rodriguez, H; Curiel, JA; Landete, JM; de las Rivas, B; López de Felipe, F; Gómez-Cordovés, C; Mancheño, JM; Muñoz, R International Journal of Food Microbiology (2009) 132, 79-90 (doi:10.1016/j.ijfoodmicro.2009.03.025) Rodríguez, H; Landete, JM; Curiel, JA; de Las Rivas, B; Mancheño, JM; Muñoz, R J. Agric. Food Chem. (2008) 56(9), 3068-3072 (doi:10.1021/jf703779s) Rodríguez, H; de las Rivas, B; Muñoz, R; Mancheño, JM Acta Crystallographica (2007) F63, 300-303 (doi:10.1107/S1744309107008846) |
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Catabolic ornithine transcarbamylase from
Lactobacillus hilgardii Ornithine transcarbamylases (OTCs) are oligomeric enzymes involved in the metabolism of the amino acid arginine. The structural analyses of catabolic OTC from Lactobacillus hilgardii have revealed a new oligomeric state (hexamer) within the family of ornithine transcarbamylases, and also the presence of a metal binding-site. Selected references: de Las Rivas, B; Fox, GC; Angulo, I; Martinez-Ripoll, M; Rodríguez, H; Muñoz, R; Mancheño, JM Journal of Molecular Biology (2009) 393, 425-434 (doi:10.1016/j.jmb.2009.08.002) de Las Rivas, B; Rodríguez, H; Angulo, I; Muñoz, R; Mancheño, JM Acta Crystallographica (2007) F63, 563-567 (doi:10.1107/S1744309107025195) |
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Structural and Biotechnological Characterization of Lipases Project Leader: Juan A. Hermoso |
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| The lipolytic enzymes belong to a large
family of enzymes that facilitate the degradation of lipids. Lipases
are members of this family that we have investigated extensively.
Lipolytic enzymes are water-soluble enzymes that are characterized by
their ability to hydrolize aggregated lipids with a much higher
velocity than the same lipid in its monomolecular form. Our work aims
structural characterization of different lipases, their activation
mechanism and their biotechnological applications. |
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Activation Mechanism of Bacterial Thermoalkalophilic Lipases The bacterial thermoalkalophilic lipases hydrolyze saturated fatty acids at 60–75 °C and pH 8–10. The crystal structure of the lipase from Geobacillus thermocatenulatus, shows that enzyme activation involves large structural rearrangements of around 70 amino acids and the concerted movement of two lids. The movements are stabilized by a Zn2+ binding domain, which is characteristic of this family of lipases. Two detergent molecules are placed in the active site, mimicking chains of the triglyceride substrate, demonstrating the position of the oxyanion hole and the three pockets that accommodate the sn-1, sn-2 and sn-3 fatty acids chains. The combination of structural and biochemical studies indicate that the lids opening is not mediated by temperature but triggered by interaction with lipid substrate. This opening implies dramatic structural rearrangements that can be seen through this link. Selected references: Carrasco-López, C; Godoy, C; de las Rivas, B; Fernandez-Lorente, G; Palomo, JM; Guisán, JM; Fernández-Lafuente, R; Martínez-Ripoll, M; Hermoso, JA Journal of Biological Chemistry (2009) 284, 4365-4372 (doi:10.1074/jbc.M808268200) |
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Biotechnological Applications of Lipases Selected references: Fuciños, P; Pastrana, L; Sanromán, A; Longo, MA; Hermoso, JA; Rúa ML Journal of Molecular Catalysis B: Enzymatic (2011) 70, 127-137 (doi:10.1016/j.molcatb.2011.02.017) Di Lorenzo, M; Hidalgo, A; Molina, R; Hermoso, JA; Greco, G; Bornscheuer, UT Applied and Environmental Microbiology. (2007) 73(22), 7291-7299 Palomo, JM; Fernández-Lorente, G; Ortiz, C; Segura, RL; Mateo, C; Fuentes, M; Hermoso, JA; Fernández-Lafuente, R; Guisán, JM Medicinal Chemistry Reviews-Online (2005) 2(5), 369-378 (doi:10.2174/156720305774330494) |
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Degradation Mechanism of Plant Cell Walls Enzymes As a component of the array of enzymes produced by micro-organisms to deconstruct plant cell walls, feruloyl esterases hydrolyze phenolic groups involved in the cross-linking of arabinoxylan to other polymeric structures. This is important for opening up the cell wall structure, making material more accesible to glycosyl hydrolases. Here we describe the first crystal structure of the non-modular type-A feruloyl esterase from Aspergillus niger (AnFAeA) solved at 2.5 A resolution. AnFaeA displays an alpha/beta hydrolase fold similar that found in fungal lipases and different to that reported for other feruloyl esterases. Selected references: Faulds, C; Molina, R; Gonzalez, R; Husband, F; Juge, N; Sanz-Aparicio, J; Hermoso, JA FEBS Journal. (2005), 272, 4362-4371 Hermoso, JA; Sanz-Aparicio, J; Molina, R; Juge, N; Gonzalez, R; Faulds, C Journal of Molecular Biology (2004) 338, 495-506 |
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Activation Mechanism of Candida rugosa Lipases The crystal structure of the closed state of the Candida rugosa lipase 2 has permited us to achieve a comprehensive structural comparison among members of this family of enzymes, which has yielded clues defining their lipase/esterase character. Selected references: Otero, C; Fernández, M; Hermoso, JA; Martínez-Ripoll, M Journal of Molecular Catalysis B: Enzymatic, (2005). 32, 225-229 Mancheño, JM; Pernas, MA; Martínez, MJ; Ochoa, B; Rúa, ML; and Hermoso, JA Acta Crystallographica (2003) D59, 499-501 Mancheño, JM; Pernas, MA; Martínez, MJ; Ochoa, B; Rúa, ML; Hermoso, JA Journal of Molecular Biology (2003) 332, 1059-1069 |
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Activation
Mechanism and Hydrolitic Machinery of Pancreatic Lipases Enzymes, such as Pancreatic Lipases, are the key enzymes in the absorption of dietary fats. The three dimensional structure of the Lipase-Colipase complex, pictured here, and the neutron crystal structure of the Lipase-Colipase-Micelle complex revealed new insights on the activation mechanisms of these enzymes. Selected references: Pignol, D; Ayvazian, L; Kerfelec, B; Timmins, P; Crenon, I; Hermoso, JA; Fontecilla-Camps, J; Chapus, C Journal of Biological Chemistry (2000) 275, 4220-4224 [Abstract] [PDF-copy] Pignol, D; Hermoso, JA; Kerfelec, B; Crenon, I; Chapus, C; Fontecilla-Camps, J Chem. and Phys. of Lipids (1998), 93, 123-129 [Abstract] Ayvazian, L; Crenon, I; Hermoso, JA; Pignol, D; Chapus, C; Kerfelec, B Journal of Biological Chemistry (1998) 273, 33604-33609 [Abstract] [PDF-copy] Crenon, I; Jayne, S; Kerfelec, B; Hermoso, JA; Pignol, D; Chapus, C Biochemical and Biophysical Research Communications (1998) 246(2), 513-517 [Abstract] Crenon, I; Flogizzo, E; Kerfelec, B; Virine, A; Pignol, D; Hermoso, JA; Bonicel, J; Chapus, C Protein Engineering (1998) 11, 135-142 [Abstract] Hermoso, JA; Pignol, D; Penel, S; Roth, M; Chapus, C; Fontecilla-Camps, J EMBO Journal (1997) 16, 5531-553 [Abstract] [PDF-copy] Hermoso, JA; Pignol, D; Kerfelec, B; Crenon, I; Chapus, C; Fontecilla-Camps, J Journal of Biological Chemistry (1996) 271, 18007-18016 [Abstract] [PDF-copy] [Further Information] |
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Electron Transfer Proteins Project Leader: Juan A. Hermoso |
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Insights into Sequential Catalysis of FAD Synthetase in Prokaryotes The crystal structure of the modular FAD synthetase (FADS) has been solved at 1.95 Å resolution. The FADS structure presents two catalytic modules, a C-terminal with ATP:riboflavinkinase activity and a N-terminal with ATP:FMN adenylyltransferase activity, responsible of the synthesis of FAD from riboflavin in two sequential steps. Crystallographic and biophysical studies revealed a hexameric assembly formed by the interaction of two trimers involved in regulation of the catalytic efficiency of the modular enzyme. Selected references: Herguedas, B; Martínez-Júlvez, M; Frago, S; Medina, M; Hermoso, JA Journal of Molecular Biology (2010) 400, 218–230 (doi:10.1016/j.jmb.2010.05.018) |
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Electron Transfer and Catalytic Mechanisms in FPR reductase and NifF flavodoxin from Rhodobacter capsulatus Selected references: Goñi, G.; Herguedas, B; Hervás, M.; Peregrina, JR; De la Rosa, MA; Gómez-Moreno, C; Navarro, JA; Hermoso, JA; Martínez-Júlvez, M; Medina, M Biochimica et Biophysica Acta (2009) 1787, 144–154 (doi:10.1016/j.bbabio.2008.12.006) Pérez-Dorado, I; Hermoso, JA; Goñi, G; Medina, M; Bortolotti, A; Carrillo, N; Cortez, N in Flavins and Flavoproteins (2008), Frago, S; Gómez-Moreno, C; Medina, M; Eds., pp. 267-273, ISBN: 978-84-7733-017-2 Pérez-Dorado, I; Bortolotti, A; Cortez, N; Hermoso, JA in Flavins and Flavoproteins (2008). Frago, S; Gómez-Moreno, C; Medina, M; Eds., pp. 261-267, ISBN: 978-84-7733-017-2 Pérez-Dorado, I; Bortolotti, A; Cortez, N; Hermoso, JA Acta Crystallographica (2008) F64, 375-377 Nogués, I; Pérez-Dorado, I; Frago, S; Bittel, C; Mayhew, SG; Gómez-Moreno, C; Hermoso, JA; Medina, M; Cortez, N; Carrillo, N Biochemistry (2005) 44, 11730-11740 Pérez-Dorado, I; Bittel, C; Cortez, N; Hermoso, JA Acta Crystallographica (2004) D60, 2332-2335 |
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Cofactor Binding in Helicobacter pylori Flavodoxin Selected references: Martínez-Júlvez, M; Cremades, N; Bueno, M; Pérez-Dorado, I; Maya, C; Cuesta-López, S; Prada, D; Falo, F; Hermoso, JA; Sancho, J Proteins: Structure, Function and Bioinformatics (2007) 69, 581-594 |
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FMN binding and Oxido-Reduction properties in Anabaena Flavodoxin Selected references: Frago, S; Goñi, G; Herguedas, B; Peregrina, JR; Serrano, A; Perez-Dorado, I; Molina, R; Gómez-Moreno, C; Hermoso, JA; Martínez-Júlvez, M; Mayhew, SG; Medina, M Archives of Biochemistry and Biophysics (2007) 467, 206-217 |
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Extrinsic Proteins of Photosystem II We report the high resolution structure of the spinach PsbQ protein, one of the main extrinsic proteins of higher plant photosystem II (PSII). The crystal structure shows that there are two well defined regions in PSbQ, the C-terminal region (residues 46-149) folded as a four-helical up-down bundle and the N-terminal region (residues 1-45) that is losely packed. This structure provides, for the first time, insights into the crucial N-terminal region. First, two parallel beta-strands cross spatially, joining the beginning and the end of the N-terminal region of PsbQ. Secondly, the residues Pro9-Pro10-Pro11-Pro12 form a left-handed helix (or a polyproline type-II, PPII, structure), which is stabilized by H-bonds between the Pro peptide carbonyls and solvent water molecules. And thirdly, residues 14-33 are not visible in the electron density map, suggesting that this loop might be very flexible and presumably extended when PsbQ is free in solution. Based on the essential role of the N-terminal region of PsbQ in binding to PSII, we propose that both the PPII structure and the missing loop are key secondary structure elements in the recognition of specific protein-protein interactions between PsbQ and other oxygen-evolving complex extrinsic and/or intrinsic proteins of PSII. In addition, the PsbQ crystal coordinates two Zn+2 ions, one of them is proposed to have physiological role in higher plants based on the fully conservation of the ligand protein residues in the sequence subfamily. Selected references: Balsera, M; Arellano, JB; Revuelta, JL; de las Rivas, J; Hermoso, JA Journal of Molecular Biology (2005) 350, 1051-1060 |
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Electron Transfer and Catalytic Mechanisms in Anabaena FNR Ferredoxin NADP+ reductase (FNR) catalyzes the final electron transport step of linear photosynthesis. In photosynthesis, the energy of an absorbed photon is transferred in the form of electrons through a series of electron carriers ultimately forming NADPH. The three dimensional structure of differents mutants of FNR, and their complexes with the electron carriers and with inhibitors are currently on going. Selected references: Peregrina, JR; Herguedas, B; Hermoso, JA; Martínez-Júlvez, M; Medina, M Biochemistry (2009) 48, 3109-3119 Tejero, J; Pérez-Dorado, I; Maya, C; Martínez-Júlvez, M; Sanz-Aparicio, J; Gómez-Moreno, C; Hermoso, JA; Medina, M Biochemistry (2005).44, 13477-13490 Mayoral, T; Martínez-Júlvez, M; Pérez-Dorado, I; Sanz-Aparicio, J; Gómez-Moreno, C; Medina, M; Hermoso, JA Proteins: Structure, Function and Bioinformatics (2005) 59, 592-602 Hermoso, JA; Mayoral, T; Faro, M; Gomez-Moreno, C; Sanz-Aparicio, J; Medina, M Journal of Molecular Biology (2002), 319, 1193-1142 Faro, M; Frago, S; Mayoral, T; Hermoso, JA; Sanz-Aparicio, J; Gómez-Moreno, C; Medina, M European Journal of Biochemistry (2002) 269, 4938-4947 Medina, M; Luquita, A; Tejero, J; Hermoso, JA; Mayoral, T; Sanz-Aparicio, J; Grever, K; Gomez-Moreno, C Journal of Biological Chemistry (2001), 276, 11902-11912 Martínez-Júlvez, M; Nogués, I; Faro, M; Hurley, JK; Brodie, TB; Mayoral, T; Sanz-Aparicio, J; Hermoso, JA; Stankovich, MT; Medina, M; Tollin, G; Gómez-Moreno, C Journal of Biological Chemistry (2001) 276, 27498-27510 Mayoral, T; Medina, M; Sanz-Aparicio, J; Gomez-Moreno, C; Hermoso, JA Proteins: Structure, Function, and Genetics (2000), 38, 60-69 Martinez-Julvez, M; Hermoso, JA; Hurley, JK; Mayoral, T; Sanz-Aparicio, J; Tollin, G; Gomez-Moreno, C; Medina, M Biochemistry (1998), 37, 17680-17691 |
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Structure and Regulation of Methionine Cycle Enzymes Project Leader: Julia Sanz-Aparicio |
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| The methionine cycle involves a series of reactions that are essential in cellular metabolism. It regulates between the amino acids methionine and cysteine for protein synthesis and supplies the substrate for polyamine syntesis. Moreover, it provides the biochemicals that produce critical components of the methylation reaction and, therefore, it is implicated within a wide range of functions including protein production, DNA regulation and neurotransmitter production. Abnormalities in methionine metabolism are associated with cardiovascular and liver disease, neural tube defects, and cancer, and due to its essential role in the cell, it has been the subject of many experimental studies. These studies have revealed a high complexity of the cycle, in part due to the fact that the involved enzymes are activated and inhibited by intermediates of the cycle. Our aim is to get a deeper knowledge on the enzymatic mechanisms at the molecular level in order to better understand the function and the complex regulation of the methionine cycle. | |
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Enzymatic Mechanism of Methionine Adenosyltransferase MAT enzymes are cytosolic proteins that use methionine and ATP to synthesize S-adenosylmethionine (SAM), the main methyl-group donor for the great variety of transmethylation reactions in all organisms. Alterations in SAM levels have been detected in several diseases and hence the control of MAT activity has proved to be a potent strategy in drug design. The structural studies on tetrameric MAT from rat liver reveals great important insights into the unusual two-steps enzymatic mechanism. Selected references: Pajares, MA; Gasset, M; Sanz-Aparicio, J; Calvete, JJ; Arrondo, JL Res. Adv. In Biological Chem. (2004) 2, 31-41 González, B; Pajares, MA; Hermoso, JA; Guillerm, D; Guillerm, G; Sanz-Aparicio, J Journal of Molecular Biology (2003) 331, 407-416 López-Vara, MC; Martínez-Chantar, ML; González, B; Gasset, M; Alvarez, L; Garrido, F; Hermoso, J; Sanz-Aparicio, J; Pajares, M in Methionine metabolism: molecular mechanisms and clinical implications (2000) Ed. Master Line, S.L. ISBN: 84-930358-5-2 González-Pérez, B; Pajares, MA; Hermoso, JA; Alvarez, L; Garrido, F; Sanz-Aparicio, J Journal of Molecular Biology (2000) 300, 363-375 |
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Structure of Betaine Homocysteine Methyltransferase Betaine-homocysteine S-methyltransferase (BHMT) is one of the two enzymes known to methylate homocysteine to produce methionine in the liver. This enzyme is a focus of great interest due to the role of homocysteine as a potential independent risk factor for cardiovascular diseases. BHMT is an oligomer composed of four identical subunits, that belong to the thiol-selenol methyltransferases family. The 3d structure shows the mechanism activating the thiol substrate which is common to the family. It also reveals the conformational changes produced during the enzymatic reaction and the structural requirements which are specific for the different functionality within the family. Selected references: Garrido, F; Gasset, M; Sanz-Aparicio, J; Alfonso, C; Pajares, MA Biochemical Journal (2005) 391, 589-599 Gonzalez-Perez, B; Pajares, MA; Martinez-Ripoll, M; Blundell, TL; Sanz-Aparicio, J Journal of Molecular Biology (2004) 338, 771-782 [PDF-copy] Gonzalez-Perez, B; Sanz-Aparicio, J; Campillo, N; Pajares, MA Biochemical Journal (2003) 370, 945-952 Gonzalez-Perez, B; Pajares, MA; Too, HP; Garrido, F; Blundell, TL; Sanz-Aparicio, J Acta Crystallographica (2002) D58, 1507-1510 |
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Pore-forming toxins Project Leader: Jose M. Mancheño |
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Combining crystallographic and functional approaches, we have analyzed at high-resolution the sugar-binding mode of the recombinant N-terminal ricin-B domain of the haemolytic protein LSLa (LSL150) from the mushroom Laetiporus sulphureus, and also provided in vitro evidences suggesting that, together with its putative receptor-binding role, this module may also increase the solubility of its membrane pore-forming partner. We firstly demonstrate that recombinant LSL150 behaves as an autonomous folding unit and an active lectin. We have determined its crystal structure at 1.47 Å resolution, and also that of the [LSL150:(lactose)β,γ] binary complex at 1.67 Å resolution. This complex reveals two lactose molecules bound to the beta and gamma sites of LSL150, respectively. Isothermal titration calorimetry indicates that LSL150 binds two lactoses in solution with highly different affinities. Also, we test the working hypothesis that LSL150 exhibits in vivo properties typical of solubility tags. With this aim, we have fused an engineered version of LSL150 (LSLt) to the N-terminal end of various recombinant proteins. All the designed LSL150–tagged fusion proteins were successfully produced at high yield and, furthermore, the target proteins were purified by a straightforward affinity procedure on agarose-based matrices due to the excellent properties of LSL150 as affinity tag. An optimized protocol for target protein purification was devised which involved removal of the LSL150 tag through in-column cleavage of the fusion proteins with His6-tagged TEV endoprotease. These results permitted to set up a novel, lectin-based system for production and purification of recombinant proteins in E. coli cells with attractive biotechnological applications. Selected references: Angulo, I; Acebrón, I; de las Rivas, B; Muñoz, R; Rodríguez-Crespo, I; Menéndez, M; García, P; Tateno, H; Goldstein, IJ; Pérez-Agote, B; Mancheño, JM Glycobiology (2011) 21, 1349-1361 (doi:10.1093/glycob/cwr074) |
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The crystal structure of the novel hexameric hemolytic lectin LSL from the parasitic mushroom Laetiporus sulphureus has revealed for the first time the presence of a pore-forming module which is shared with pore-forming toxins from bateria of the aerolysin family. Selected references: Mancheño, JM; Tateno, H; Goldstein, IJ; Martínez-Ripoll, M; Hermoso, JA Journal of Biological Chemistry (2005) 280, 17251-17259 [PDF-copy] Mancheño, JM; Hiroaki, T; Goldstein, IJ; Hermoso, JA Acta Crystallographica (2004) D60, 1139-1141 |
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The
high resolution crystal structures of the free and phosphocholine-bound
water-soluble state of the actinoporin Sticholysin II, and that of a
tetrameric oligomer in a lipidic interface obtained by electron
microscopy have provided insights into the molecular mechanism of
membrane pore formation. Selected references: Mancheño, JM; Martín-Benito, J; Martínez-Ripoll, M; Gavilanes, JG; Hermoso, JA Structure (2003) 11, 1319-1328 [PDF-copy] Mancheño, JM; Martínez-Ripoll, M; Gavilanes, JG and Hermoso, JA Acta Crystallographica (2002) D58, 1229-1231 [PDF-copy] [Further Information] |
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DNA binding proteins Project Leader: Armando Albert |
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Replication
is the process by which organisms are able to produce an identical copy
of its genetic material. Proteins responsible of replication are
arranged into a supramolecular assembly called replicative complex.
These structures are DNA production factories anchored to the cell
membrane. The X-ray and solution structures of P16.7 provide the
molecular basis of the anchorage to the membrane and the structure of
these molecular factories. Selected references: Asensio, JL; Albert, A; Muñoz-Espin, D; Gonzalez, C; Hermoso, JA; Villar, L; Jimenez-Barbero, J; Salas, M; Meijer, W Journal of Biological Chemistry (2005), 21, 20730-20739 |
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Catalytic Mechanisms in Enzymes Project Leader: Armando Albert |
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Aspartate
decarboxylase overlaps, in the same active site, activities for self
procesing after transduction, synthesis of an essential cofactor
(pyruvoil group) and decarboxylation of L-aspartate. Selected references: Castillo, RM; Mizuguchi, K; Dhanaraj, V; Albert, A; Blundell, TL; Murzin, AG Structure (1999) 7, 227-236 Albert, A; Dhanaraj, V; Genshel, U; Khan, G; Ramjee, MK; Pulido, R; Sibanda, BL; Delft, F; Witty, M; Blundell, TL; Smith, AG; Abell, C Nature Structural Biology (1998) 5, 289-293 |
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2012 Structural and kinetic insights reveal that the amino acid pair GLN228/ASN254 modulates the transfructosylating specificity of Schwanniomyces occidentalis β-fructofuranosidase, an enzyme that produces prebiotics Miguel Alvaro-Benito, M. Angela Sainz-Polo, David Gonzalez-Perez, Beatriz Gonzalez, Francisco J. Plou, Maria Fernandez-Lobato, and Julia Sanz Aparicio Journal of Biological Chemistry (2012) in press (doi:10.1074/jbc.M112.355503) An analysis of subdomain orientation, conformational change and disorder in relation to crystal packing of aspartic proteinases D. Bailey, E.P. Carpenter, A. Coker, S. Coker, J. Read, A.T., Jones, P. Erskine, C.F. Aguilar, M. Badasso, L. Toldo, F., Rippmann, J. Sanz-Aparicio, A. Albert, T.L. Blundell, N.B., Roberts, S.P. Wood and J.B. Cooper Acta Crystallographica (2012) D68, 541-552 (doi: 10.1107/S0907444912004817) Sagrario Martinez-Carrera (1925-2011) Martinez-Ripoll, M. Acta Crystallographica (2012) B68, 213–214 (doi:10.1107/S0108768112003540) Structural basis of specificity in tetrameric Kluyveromyces lactis β-galactosidase Pereira-Rodríguez, A; Fernández-Leiro, R; González-Siso, MI; Esperanza Cerdán, M; Becerra, M; Sanz-Aparicio, J Journal of Structural Biology (2012) 177, 392–401 (doi:10.1016/j.jsb.2011.11.031) Promiscuous enantioselective (–)-γ-lactamase activity in the Pseudomonas fluorescens esterase I Leticia Luciana Torres, Marlen Schmidt, Anna Schliessmann, Noella Silva-Martin, Juan Hermoso, José Berenguer, Uwe Bornscheuer and Aurelio Hidalgo Organic & Biomolecular Chemistry (2012) 10, 3388-3392 (doi: 10.1039/C2OB06887G) 2011 Nobel de Química 2011 a Daniel Shechtman por su descubrimiento de los cuasicristales y de una simetría "imposible" Julia Sanz-Aparicio mi+d (2011) published in Análisis Madri+d on 15 Nov. 2011 [PDF-copy] A Practical Two-Step Synthesis of Imidazo[1,2-a]pyridines from N-(prop-2-yn-1-yl)pyridin-2-amines D. Sucunza, A. Samadi, M. Chioua, D.B. Silva, C. Yunta, L. Infantes, M.C. Carreiras, E. Soriano, J. Marco-Contelles Chemical Communications (2011) 47, 5043-5045 (doi:10.1039/C1CC10641D) Quaternary α,α,-2-Oxoazepane alpha-Amino Acids: Synthesis from Ornithine-Derived β-Lactams and Incorporation into Model Dipeptides D. Nuñez-Villanueva, M.A. Bonache, L. Infantes, M.T. Garcia-Lopez, M. Martin-Martinez, R. Gonzalez-Muniz Journal of Organic Chemistry (2011) 76, 6592-6603 (doi:10.1021/jo200894d) Crystallization and preliminary crystallographic analysis of a C2 protein from Arabidopsis thaliana Maira Diaz, Lesia Rodriguez, Miguel Gonzalez-Guzman, Martin Martinez-Ripoll and Armando Albert Acta Crystallograhica (2011) F67, 1575–1578 (doi:10.1107/S1744309111040541) Preliminary X-ray analysis of twinned crystals of the Q88Y25_Lacpl esterase from Lactobacillus plantarum WCFS1 Yanaisis Alvarez, Maria Esteban-Torres, Ivan Acebron, Blanca de las Rivas, Rosario Muñoz, Martin Martinez-Ripoll and Jose M. Mancheño Acta Crystallographica (2011) F67, 1436–1439 (doi:10.1107/S1744309111036682) The structure of Arabidopsis thaliana OST1 provides insights into the kinase regulation mechanism in response to osmotic stress Cristina Yunta, Martín Martínez-Ripoll, Jian-Kang Zhu and Armando Albert Journal of Molecular Biology (2011) 414, 135–144 (doi:10.1016/j.jmb.2011.09.041) Fructo-oligosaccharide synthesis by mutant versions of Saccharomyces cerevisiae invertase Lafraya, A., Sanz-Aparicio, J., Polaina, J. and Marín-Navarro, J. Applied Environmental Microbiology (2011) 77, 6148-6157 (doi:10.1128/AEM.05032-11) Crystallization and Preliminary X-Ray Diffraction Analysis of phosphoglycerate kinase from Streptococcus pneumoniae N. Bernardo-García, S.G. Bartual, M. Fulde, S. Bergmann and J.A. Hermoso Acta Crystallographica (2011) F67, 1285-1289 (doi:10.1107/S1744309111030922) Crystallization and Preliminary X-Ray Diffraction Analysis of the transcriptional repressor Paax, the main regulator of the phenylacetic acid degradation pathway in Escherichia coli W. A. Rojas-Altuve, C. Carrasco-López, V.M. Hernández-Rocamora, J.M. Sanz and J.A. Hermoso Acta Crystallographica (2011) F67, 1278-1280 (doi:10.1107/S1744309111029873) Crystal structures of bacterial peptidoglycan amidase ampd and an unprecedented activation mechanism Cesar Carrasco-Lopez, Alzoray Rojas-Altuve, Welie Zhang, Dusan Hesek, Mijoon Lee, Sophie Barbe, Isabelle André, Pilar Ferrer, Noella Silva-Martin, Germán R. Castro, Martin Martinez-Ripoll, Shahriar Mobashery, and Juan A. Hermoso Journal of Biological Chemistry (2011) 286, 31714-31722 (doi:10.1074/jbc.M111.264366) High-resolution structural insights on the sugar-recognition and fusion tag properties of a versatile β-trefoil lectin domain from the mushroom Laetiporus sulphureus Angulo, I., Acebrón, I., de las Rivas, B., Muñoz, R., Rodríguez-Crespo, I., Menéndez, M., García, P., Tateno, H., Goldstein, I.J., Pérez-Agote, B. and Mancheño, J.M. Glycobiology (2011) 21, 1349-1361 (doi:10.1093/glycob/cwr074) An esterase from Thermus thermophilus HB27 with hyper-thermoalkalophilic properties: purification, characterisation and structural modelling Fuciños, P., Pastrana, L., Sanromán, A., Longo, M.A., Hermoso, J.A., Rúa M.L. Journal of Molecular Catalysis B: Enzymatic (2011) 70, 127-137 (doi:10.1016/j.molcatb.2011.02.017) High-Resolution Crystal Structure of an Outer Membrane-Anchored Endolytic Peptidoglycan Lytic Transglycosylase (MltE) from Escherichia coli Cecilia Artola-Recolons, Cesar Carrasco-López, Leticia Irene Llarrull, Malika Kumarasiri, Elena Lastochkin, Iñaki Martínez de Ilarduya, Kathrin Meindl, Isabel Usón, Shahriar Mobashery and Juan A. Hermoso Biochemistry (2011) 50, 2384-2386 (doi:10.1021/bi200085y) A self assembled 3-D network propagated by coordination polymerization and H-bonding: synthesis and X-ray crystal structure of [{Co(L)2(H2O)2}(ClO4)2(CH3COCH3)2(H2O)2]n , where L = N,N-diisopropylisonicotinamide Ajay Pal Singh Pannu, Pratibha Kapoor, Geeta Hundal, Ramesh Kapoor, Martin Martinez-Ripoll and Maninder Singh Hundal Journal of Coordination Chemistry (2011) 64, 1566-1577 (doi:10.1080/00958972.2011.574286) SnRK2.6/OST1 from Arabidopsis thaliana: cloning, expression, purification, crystallization and preliminary X-ray analysis of K50N and D160A mutants Cristina Yunta, Martin Martinez-Ripoll and Armando Albert Acta Crystallographica (2011) F67, 364–368 (doi:10.1107/S1744309110053807) Crystallization and Preliminary X-Ray Diffraction Analysis of Lytic Transglycosylase MltE from Escherichia coli Cecilia Artola-Recolons, Leticia Llarrull, Elena Lastochkin, Shahriar Mobashery and Juan A. Hermoso Acta Crystallographica (2011) F67, 161-163 (doi:10.1107/S1744309110049171) New Alkaloid Antibiotics that Target the DNA Topoisomerase I of Streptococcus pneumoniae Teresa García, María Amparo Blázquez, María José Ferrándiz, María Jesús Sanz, Noella Silva-Martín, Juan A. Hermoso and Adela G. de la Campa Journal of Biological Chemistry (2011) 286, 6402-6413 (doi:10.1074/jbc.M110.148148) The pURI family of expression vectors: A versatile set of ligation independent cloning plasmids for producing recombinant His-fusion proteins. Curiel, J.A., de Las Rivas., B, Mancheño, J.M., Muñoz, R. Protein Expression and Purification (2011) 76, 44-53 (doi:10.1016/j.pep.2010.10.013) 2010 Crystallographers in Spain Martín Martínez-Ripoll IUCr Newsletter (2010) 18(4), 5-13 [PDF-copy] Crystallography in Spain Martín Martínez-Ripoll IUCr Newsletter (2010) 18(3), 5-9 [PDF-copy] Cristalografía en España Martín Martínez-Ripoll Anales de Química (2010) 106, 319-329 [PDF-copy] Biochemical characterization of the transcriptional regulator BzdR from Azoarcus sp. CIB Durante-Rodriguez, G., Valderrama, A., Mancheño, J.M., Rivas, G., Alfonso, C., Arias-Palomo, E., Llorca, O., Garcia, J.L., Diaz, E., Carmona, M. Journal of Biological Chemistry (2010) 285, 35694-35705 (doi:10.1074/jbc.M110.143503) Synthesis of (E)-diethyl 6,60-(diazene-1,2-diyl)bis(5-cyano-2-methyl-4-phenylnicotinates), a new type of 2,20-azopyridine dyesodification of pancreatic lipase properties by directed molecular evolution Botelho da Silva, D., Samadi, A., Infantes, L., Carreiras, M.C. and Marco-Contelles, J. Tetrahedron Letters (2010) 51, 6278-6281 (doi:10.1016/j.tetlet.2010.09.095) Modification of pancreatic lipase properties by directed molecular evolution Colin, D.Y., Deprez-Beauclair, P., Silva, N., Infantes, L. and Kerfelec, B. Protein Engineering Design & Selection (2010) 23, 365-373 (doi:10.1093/protein/gzq008) New insights into the fructosyltransferase activity of Schwanniomyces occidentalis β-fructofuranosidase, emerging from nonconventional codon usage and directed mutation Alvaro-Benito, M., Abreu, M., Portillo, F., Sanz-Aparicio, J. and Fernández-Lobato, M. Applied Environmental Microbiology (2010) 76, 7491-7499 (doi:10.1128/AEM.01614-10) Crystallization and preliminary X-ray diffraction analysis of the fructofuranosidase from Xanthophyllomyces dendrorhous Polo, A., Linde, D., Estévez, M., Fernández-Lobato, M. and Sanz-Aparicio, J. Acta Crystallographica (2010) F66, 1441-1444 (doi:10.1107/S1744309110029192) Crystallization and preliminary X-ray diffraction analysis of Pseudomonas aeruginosa phosphorylcholine phosphatase L.H. Otero, P.R. Beassoni, C.E. Domenech, A.T. Lisa and A. Albert Acta Crystallographica (2010) F66, 957-960 (doi:10.1107/S1744309110024061) Structural analysis of Saccharomyces cerevisiae α-galactosidase and its complexes with natural substrates reveals new insights into substrate specificity of GH27 glycosidases Rafael Fernández-Leiro, Ángel Pereira-Rodríguez, M. Esperanza Cerdán, Manuel Becerra and Juliana Sanz-Aparicio Journal of Biological Chemistry (2010) 285, 28020-28033 (doi:10.1074/jbc.M110.144584) Inositol 1,3,4,5,6-pentakisphosphate 2-kinase is a distant IPK member with a singular inositide binding site for axial 2-OH recognition B. González, J. Baños-Sanz, M. Villate, C. Brearley and J. Sanz-Aparicio Proceedings of the National Academy of Sciences (2010) 107, 9608-9613 (doi:10.1073/pnas.0912979107) Insights into pneumococcal fratricide from crystal structure of the modular Killing Factor LytC Inmaculada Pérez-Dorado, Ana González, María Morales, Reyes Sanles, Waldemar Striker, Waldemar Vollmer, Shahriar Mobashery, José L. García, Martín Martínez-Ripoll, Pedro García and Juan A. Hermoso Nature Structural & Molecular Biology (2010) 17, 576-582 (doi:10.1038/nsmb.1817) Promotion of multipoint covalent immobilization through different regions of genetically modified penicillin G acylase from E. coli Grazú V., López-Gallego F., Montes T., Abian O., González R., Hermoso J.A., García J.L., Mateo C. and Guisán J.M. Process Biochemistry (2010) 45, 390-398 (doi:10.1016/j.procbio.2009.10.013) Pneumococcal CbpD is a murein hydrolase that requires a dual cell envelope binding specificity to kill target cells during fratricide Vegard Eldholm, Ola Johnsborg, Daniel Straume, Hilde Solheim Ohnstad, Kari Helene Berg, Juan A. Hermoso and Leiv Sigve Håvarstein Molecular Microbiology (2010) 76, 905-917 (doi:10.1111/j.1365-2958.2010.07143.x) Crystallization and preliminary crystallographic analysis of the catalytic module of endolysin from Cp-7, a phage infecting Streptococcus pneumoniae Noella Silva-Martin, Rafael Molina, Ivan Angulo, José M. Mancheño, Pedro García and Juan A. Hermoso Acta Crystallographica (2010) F66, 670–673 (doi:10.1107/S1744309110006718) Crystallization of the pneumococcal autolysin LytC: in-house phasing using novel lanthanide complexes I. Pérez-Dorado, R. Sanles, A. González, P. García, J.L. García, M. Martínez-Ripoll and J.A. Hermoso Acta Crystallographica (2010) F66, 448-451 (doi:10.1107/S1744309110006081) Oligomeric State in the Crystal Structure of Modular FAD Synthetase Provides Insights into Its Sequential Catalysis in Prokaryotes Beatriz Herguedas, Marta Martínez-Júlvez, Susana Frago, Milagros Medina and Juan A. Hermoso Journal of Molecular Biology (2010) 400, 218–230 (doi:10.1016/j.jmb.2010.05.018) p-Coumaric acid decarboxylase from Lactobacillus plantarum: Structural insights into the active site and decarboxylation catalytic mechanism Héctor Rodríguez, Iván Angulo, Blanca de las Rivas, Nuria Campillo, Juan A. Páez, Rosario Muñoz and José M. Mancheño Proteins: Structure, Function, and Bioinformatics (2010) 78, 1662-1676 (doi:10.1002/prot.22684) Laetiporus sulphureus Lectin and Aerolysin Protein Family Jose Miguel Mancheño, Hiroaki Tateno, Daniel Sher and Irwin J. Goldstein in Proteins: Membrane Binding and Pore Formation (2010), Chp. 6, pp. 67-80. Gregor Anderluh and Jeremy Lakey, Eds., Landes Bioscience. ISBN: 978-1-4419-6326-0. See also Advances in Experimental Medicine and Biology (2010) 677, 67-80 Structural and kinetic analysis of Schwanniomyces occidentalis invertase reveals a new oligomerization pattern and the role of its supplementary domain in substrate binding Alvaro-Benito, M., Polo, A., González, B., Fernández-Lobato, M. and Sanz-Aparicio, J. Journal of Biological Chemistry (2010) 285, 13930-13941 (doi:10.1074/jbc.M109.095430) Structure of GroEL in Complex with an Early Folding Intermediate of Alanine Glyoxylate Aminotransferase Armando Albert, Cristina Yunta, Rocío Arranz, Álvaro Peña, Eduardo Salido, José María Valpuesta and Jaime Martín-Benito Journal of Biological Chemistry (2010) 285, 6371-6376 (doi:10.1074/jbc.M109.062471) Crystallization and preliminary X-ray crystallographic analysis of β-galactosidase from Kluyveromyces lactis Pereira-Rodríguez, A. Fernández-Leiro, R., González-Siso, M., Cerdán, E., Becerra, M. and Sanz-Aparicio, J. Acta Crystallographica (2010) F66, 297-300 (doi:10.1107/S1744309109054931) Crystallization and preliminary X-ray diffraction analysis of inositol 1,3,4,5,6-pentakisphosphate kinase from Arabidopsis thaliana Baños-Sanz, J.I., Villate, M., Sanz-Aparicio, J., Brearley, C.A. and González B. Acta Crystallographica (2010) F66, 102-106 (doi:10.1107/S1744309109051057) Crystallization and preliminary X-ray diffraction analysis of α-galactosidase from Saccharomyces cerevisiae Fernández-Leiro, F., Pereira, A., Cedán, E. Becerra-Fernández, M. and Sanz-Aparicio, J. Acta Crystallographica (2010) F66, 44-47 (doi:10.1107/S1744309109047794) Structural insights into the specificity of Xyn10B from Paenibacillus barcinonensis and its improved stability by forced protein evolution Gallardo, O. Pastor, F.J., Polaina, J., Díaz, P., Vogel, P. Isorna, P., González-Pérez, B. and Sanz-Aparicio, J. Journal of Biological Chemistry (2010) 285, 2721-2733 (doi:10.1074/jbc.M109.064394) 2009 Sticholysins, two pore-forming toxins produced by the Caribbean Sea anemone Stichodactyla helianthus: their interaction with membranes C. Alvarez, J.M. Mancheño, D. Martínez, M. Tejuca, F. Pazos and M.E. Lanio Toxicon (2009) 54, 1135-1147 (doi:10.1016/j.toxicon.2009.02.022) Food phenolics and lactic acid bacteria H. Rodriguez, J.A. Curiel, J.M. Landete, B. de las Rivas, F. López de Felipe, C. Gómez-Cordovés, J.M. Mancheño and R. Muñoz International Journal of Food Microbiology (2009) 132, 79-90 (doi:10.1016/j.ijfoodmicro.2009.03.025) Production and physicochemical properties of recombinant Lactobacillus plantarum tannase J.A. Curiel, H. Rodríguez, I. Acebrón, J.M. Mancheño, B. de Las Rivas and R. Muñoz Journal of Agricultural and Food Chemistry (2009) 57, 6224-6230 (doi:10.1021/jf901045s) Crystal structure of the hexameric catabolic ornithine transcarbamylase from Lactobacillus hilgardii: Structural insights into the oligomeric assembly and metal binding B. de Las Rivas, G.C. Fox, I. Angulo, M. Martinez-Ripoll, H. Rodríguez, R. Muñoz and J.M. Mancheño Journal of Molecular Biology (2009) 393, 425-434 (doi:10.1016/j.jmb.2009.08.002) Crystallization and preliminary X-ray diffraction studies of the FAD synthetase from Corynnebacterium ammoniagenes Beatriz Herguedas, Marta Martínez-Júlvez, Milagros Medina and Juan A. Hermoso Acta Crystallographica (2009) F65, 1285-1288 (doi:10.1107/S1744309109044789) Crystallization and preliminary X-ray diffraction studies of the carbohydrate recognition domain of SIGN-R1, a receptor for microbial polysaccharides and sialylated antibody on splenic marginal zone macrophages N. Silva-Martin, J.D. Schauer, C.G. Park and J.A. Hermoso Acta Crystallographica (2009) F65, 1264-1266 (doi:10.1107/S1744309109041992) Crystallization and preliminary X-ray diffraction analysis of the fructofuranosidase from Schwanniomyces occidentalis Polo-Rivas, A. Alvaro-Benito, A., Fernández-Lobato, M. and Sanz-Aparicio, J. Acta Crystallographica (2009) F65, 1162-1165 (doi:10.1107/S174430910903938) Discovery of specific flavodoxin inhibitors as potential therapeutic agents against Helicobacter pylori infection Nunilo Cremades, Adrián Velázquez-Campoy, Marta Martínez-Júlvez, José L. Neira, Inmaculada Pérez-Dorado, Juan Hermoso, Pilar Jiménez, Angel Lanas, Paul S. Hoffman and Javier Sancho ACS Chemical Biology (2009) 4, 928-938 (doi:10.1021/cb900166q) Cloning, production, purification and preliminary crystallographic analysis of a glycosidase from the food lactic acid bacterium Lactobacillus plantarum CECT 748T Iván Acebrón, José A. Curiel, Blanca de las Rivas, Rosario Muñoz, José M. Mancheño Protein Expression and Purification (2009) 68, 177-182 (doi:10.1016/j.pep.2009.07.006) Characterization of gadolinium complexes for SAD phasing in macromolecular crystallography: application to CbpF Rafael Molina, Meike Stelter, Richard Kahn and Juan A. Hermoso Acta Crystallographica (2009) D65, 823–831 (doi:10.1107/S0907444909017958) Protein Motifs Involved in Coenzyme Interaction and Enzymatic Efficiency in Anabaena Ferredoxin–NADP+ Reductase José Ramón Peregrina, Beatriz Herguedas, Juan A. Hermoso, Marta Martínez-Júlvez and Milagros Medina Biochemistry (2009) 48, 3109-3119 (doi:10.1021/bi802077c) Crystal structure of CbpF, a bifunctional choline-binding protein and autolysis regulator from Streptococcus pneumoniae Rafael Molina, Ana González, Meike Stelter, Inmaculada Pérez-Dorado, Richard Kahn, María Morales, Susana Campuzano, Nuria E. Campillo, Shahriar Mobashery, José L. García, Pedro García and Juan A. Hermoso EMBO Reports (2009) 10, 246-251 (doi:10.1038/embor.2008.245) Activation of bacterial thermoalkalophilic lipases is spurred by dramatic structural rearrangements Cesar Carrasco-López, Cesar Godoy, Blanca de las Rivas, Gloria Fernandez-Lorente, Jose M. Palomo, José M. Guisán, Roberto Fernández-Lafuente, Martín Martínez-Ripoll and Juan A. Hermoso Journal of Biological Chemistry (2009) 284, 4365-4372 (doi:10.1074/jbc.M808268200) Coenzyme binding and hydride transfer in Rhodobacter capsulatus ferredoxin/flavodoxin NADP(H) oxidoreductase Ana Bortoloti, Inmaculada Pérez-Dorado, Guillermina Goñi, Milagros Medina, Juan A. Hermoso, Néstor Carrillo and Néstor Cortez Biochimica et Biophysica Acta - Proteins & Proteomics (2009) 1794, 199-210 (doi:10.1016/j.bbapap.2008.09.013) Flavodoxin: A compromise between efficiency and versatility in the electron transfer from Photosystem I to Ferredoxin-NADP+ reductase Guillermina Goñi, Beatriz Herguedas, Manuel Hervás, José R. Peregrina, Miguel A. de la Rosa, Carlos Gómez-Moreno, José A. Navarro, Juan A. Hermoso, Marta Martínez-Júlvez and Milagros Medina Biochimica et Biophysica Acta - Bioenergetics (2009) 1787, 144–154 (doi:10.1016/j.bbabio.2008.12.006) Further Evidence for 2-Alkyl-2-carboxyazetidines as γ-Turn Inducers J.L. Baeza, G. Gerona-Navarro, K. Thompson, M.J. Prez de Vega, L. Infantes, M.T. García-Lopez, R. Gonzalez-Muñiz and M. Martin-Martinez Journal of Organic Chemistry (2009) 74, 8203-8211 (doi:10.1021/jo901712x) Molecular Crystal Prediction Approach by Molecular Similarity: Data Mining on Molecular Aggregation Predictors and Crystal descriptors J. Fayos Crystal Growth and Design (2009) 9, 3142-3153 (doi:10.1021/cg801122m) Experimental and Theoretical Study of the Structures and Enthalpies of Formation of the Synthetic Reagents 1,3-Thiazolidine-2-thione and 1,3-Oxazolidine-2-thione M.V. Roux, M. Temprado, P. Jiménez, C, Foces-Foces, R. Notario, A.R. Parameswar, A-V. Demchenko, J.S. Chickos, C.A. Deakyne, A.K. Ludden, J-F. Liebman Journal of Physical Chemistry A (2009) 113, 10772-10778 (doi:10.1021/jp9034216) 2008 Targeted polypharmacology: discovery of dual inhibitors of tyrosine and phosphoinositide kinases Apsel B., Blair J.A., Gonzalez B., Nazif T.M., Feldman M.E., Aizenstein B., Hoffman R., Williams R.L., Shokat K.M. and Knight Z.A. Nature Chemical Biology (2008) 4, 691-699 (doi:10.1038/nchembio.117) Crystallization and preliminary X-ray diffraction studies of the BTL2 lipase from extremophilic microorganism Bacillus thermocatenulatus César Carrasco-López, César Godoy, Blanca de las Rivas, Gloria Fernández-Lorente, José M. Palomo, José M. Guisán, Roberto Fernández-Lafuente, Martín Martínez-Ripoll and Juan A. Hermoso Acta Crystallographica (2008) F64, 1043-1045 (doi:10.1107/S1744309108031928) A one-pot, simple methodology for cassette randomisation and recombination for focused directed evolution Aurelio Hidalgo, Anna Schliessmann, Rafael Molina, Juan Hermoso and Uwe T. Bornscheuer Protein Engineering, Design & Selection (2008) 21(9), 567-576 [PDF-copy] Solid-Phase Chemical Amination of a Lipase from Bacillus thermocatenulatus To Improve Its Stabilization via Covalent Immobilization on Highly Activated Glyoxyl-Agarose Gloria Fernandez-Lorente, Cesar A. Godoy, Adriano A. Mendes, Fernando Lopez-Gallego, Valeria Grazu, Blanca de las Rivas, Jose M. Palomo, Juan Hermoso, Roberto Fernandez-Lafuente, and Jose M. Guisan Biomacromolecules (2008) 9, 2553–2561 [PDF-copy] Characterization of the p-coumaric acid decarboxylase from Lactobacillus plantarum CECT 748(T) Rodríguez, H., Landete, J.M., Curiel, J.A., de Las Rivas, B., Mancheño, J.M., Muñoz, R. Journal of Agricultural and Food Chemistry (2008) 56, 3068-3072 (doi:10.1021/jf703779s) Las enzimas líticas de los bacteriófagos (Enzibióticos): Nuevas terapias contra las infecciones bacterianas Hermoso, J.A. MADRI+D (2008), Abril 29th [Web-access] Los enzibióticos progresan en un momento clave Hermoso, J.A. Diario Médico (2008) 16 mayo, 16-17 [PDF-copy] Structural characterization of a ferredoxin-NADP(H) reductase and a flavodoxin from Rhodobacter capsulatus by X-ray crystallography Inmaculada Pérez-Dorado, Ana Bortolotti, Néstor Cortez and Juan A. Hermoso in Flavins and Flavoproteins (2008). S. Frago, C. Gómez-Moreno and M. Medina Eds. (2008). 261-267. ISBN: 978-84-7733-017-2 The coenzyme binding site of bacterial ferredoxin/flavodoxin-NADP(H) reductases Inmaculada Pérez-Dorado, Juan A. Hermoso, Guillermina Goñi, Milagros Medina, Ana Bortolotti, Néstor Carrillo, and Néstor Cortez in Flavins and Flavoproteins (2008). S. Frago, C. Gómez-Moreno and M. Medina Eds. (2008). 267-273. ISBN: 978-84-7733-017-2 Crystallization and preliminary crystallographic analysis of merohedrally twinned crystals of MJ0729, a CBS-domain protein from Methanococcus jannaschii P. Fernández-Millán, D. Kortazar, M. Lucas, M.L. Martínez-Chantar, E. Astigarraga, J.A. Fernández, O. Sabas, A. Albert, J.M. Mato and L.A. Martínez-Cruz Acta Crystallographica (2008) F64, 605-609 Crystallization of a flavodoxin involved in the nitrogen fixation in Rhodobacter capsulatus I. Pérez-Dorado, A. Bortolotti, N. Cortez and J.A. Hermoso Acta Crystallographica (2008) F64, 375-377 Hydrogen-bonded layered structures in two bis(tert-butyldimethylsilyloxy)-substituted cyclic diol derivatives. Foces-Foces C and López-Rodríguez M Acta Crystallographica (2008) C64, o657-o660 Thermochemistry of 2- and 3-thiopheneacetic acids: calorimetric and computational study Temprado M, Roux MV, Jiménez P, Foces-Foces C, and Notario R The Journal of Physical Chemistry A (2008) 112, 10378-10385 Structure-energy relationship in barbituric acid: a calorimetric, computational, and crystallographic study Roux MV, Temprado M, Notario R, Foces-Foces C, Emel'yanenko VN, and Verevkin SP The Journal of Physical Chemistry A (2008) 112, 7455-7465 A case of concomitant polymorphism and spontaneous resolution: the tetragonal phase of 5-hydroxymethyl-7,7,N-trimethyl-6-oxabicyclo[3.2.1]octane-1-carboxamide Foces-Foces C, López-Rodríguez M, and Pérez C Acta Crystallographica (2008) C64, o95-o97 2007 The structure of the C-terminal domain of the protein kinase AtSOS2 bound to the calcium sensor AtSOS3 María José Sánchez-Barrena, Hiroaki Fujii, Ivan Angulo, Martín Martínez-Ripoll, Jian-Kang Zhu and Armando Albert Molecular Cell (2007) 26, 427-435 (doi:10.1016/j.molcel.2007.04.013) Taking aim on bacterial pathogens: from phage therapy to enzybiotics Juan A. Hermoso, José L García, Pedro García Current Opinion in Microbiology (2007) 10(5), 461-472 (doi:10.1016/j.mib.2007.08.002) Enhancement of the Stability of a Prolipase from Rhizopus oryzae Towards Aldehydes by Saturation Mutagenesis Mirella Di Lorenzo, Aurelio Hidalgo, Rafael Molina, Juan A. Hermoso, Guido Greco and Uwe T. Bornscheuer Applied and Environmental Microbiology (2007) 73(22), 7291-7299 Crystallization and preliminary X ray diffraction studies of the Choline Binding Protein F from Streptococcus pneumoniae Rafael Molina, Ana González, Miriam Moscoso, Pedro García, Meike Stelter, Richard Kahn and Juan A. Hermoso Acta Crystallographica (2007) F63, 742-745 (doi:10.1107/S1744309107035865) The complex between SOS3 and SOS2 regulatory domain from Arabidopsis thaliana: cloning, expression, purification, crystallization and preliminary X-ray analysis Sánchez-Barrena, M.J., Moreno-Pérez,, S., Angulo, I., Martínez-Ripoll, M. and Albert, A. Acta Crystallographica (2007) F63, 568-570 Tuning of the FMN binding and oxido-reduction properties by neighboring side chains in Anabaena Flavodoxin S. Frago, G. Goñi, B. Herguedas, J.R. Peregrina, A. Serrano, I. Perez-Dorado, R. Molina, C. Gómez-Moreno, J.A. Hermoso, M. Martínez-Júlvez, S. G. Mayhew and M. Medina Archives of Biochemistry and Biophysics (2007) 467, 206-217 Elucidation of the Molecular Recognition of Bacterial Cell Wall by Modular Pneumococcal Phage Endolysin Cpl-1 Pérez-Dorado, N.E. Campillo, B. Monterroso, D. Hesek, M. Lee, J.A. Páez, P. García, M. Martínez-Ripoll, J.L. García, S. Mobashery, M. Menéndez and J.A. Hermoso Journal of Biological Chemistry (2007) 282, 24990-24999 Common conformational changes in flavodoxins induced by FMN and anion binding: The structure of Helicobacter pylori apoflavodoxin M Martínez-Júlvez, N Cremades, M Bueno, I Pérez-Dorado, C Maya, S Cuesta-López, D Prada, F Falo, J.A. Hermoso and. J Sancho Proteins: Structure, Function and Bioinformatics (2007) 69, 581-594 Genetic Modification of the Penicillin G Acylase Surface To Improve Its Reversible Inmobilization on Ionic Exchangers Tamara Montes, Valeria Grazu, Fernando López-Gallego, Juan A. Hermoso, José L. García, Isabel Manso, Beatriz Galán, Ramón González, Roberto Fernández-Lafuente and José M. Guisán Applied and Environmental Microbiology (2007) 73(1), 312-319 Improved stabilization of genetically modified Penicillin G Acylase in the presence of organic cosolvents by co-immobilization of the enzyme with polyethylenimine Tamara Montes, Valeria Grazu, Isabel Manso, Beatriz Galán, Ramón González, Fernando López-Gallego, Juan A. Hermoso, José L. garcía, José M. Guisán and Roberto Fernández-Lafuente Adv. Synth. Catal. (2007) 349, 459-464 Insights into molecular plasticity of choline binding proteins (pneumococcal surface proteins) by SAXS Rubén M. Buey, Begoña Monterroso, Margarita Menéndez, Greg Diakun, Pablo Chacón, Juan Antonio Hermoso and J. Fernando Díaz Journal of Molecular Biology, (2007), 365 (2), 411-424 Mixed ion exchangers supports as useful ion exchangers for protein purification. Purification of penicillin G acylase from E.coli Manuel Fuentes, Pilar Batalla, Valeria Grazu, Benevides C.C. Pessela, Cesar Mateo, Tamara Montes, Juan A. Hermoso, José M. Guisán and Roberto Fernández-Lafuente Biomacromolecules (2007) 8(2), 703-707 Insights into the activation of brain serine racemase by the multi-PDZ domain glutamate receptor interacting protein, divalent cations and ATP Baumgart, F, Mancheño JM, Rodríguez-Crespo, I FEBS J. (2007) 274(17), 4561-4571 Overexpression, purification, crystallization and preliminary structural studies of catabolic ornithine transcarbamylase from Lactobacillus hilgardii de las Rivas, B, Rodríguez, H, Angulo, I, Muñoz, R, Mancheño JM Acta Crystallographica (2007) F63, 563-567 Expression vectors for enzyme restriction- and ligation-independent cloning for producing recombinant His-fusion proteins de las Rivas, B, Curiel, JM., Mancheño JM, Muñoz, R Biotechnol. Prog. (2007) 23, 680-686 Overexpression, purification, crystallization and preliminary structural studies of p-coumaric acid decarboxylase from Lactobacillus plantarum Rodríguez, H, de las Rivas, B, Muñoz, R, Mancheño, JM Acta Crystallographica (2007) F63, 300-303 Crystal structures of Paenibacillus polymyxa β-glucosidase B complexes reveal the molecular basis of substrate specificity and give insights into the catalytic mechanism of family I glycosidases Isorna, P., Polaina, J., Latorre-García. L., Cañada, F.J., González, B. y Sanz-Aparicio, J. Journal of Molecular Biology (2007) 371, 1204-1218 Xylanases: Molecular properties and applications Pastor, F.J., Gallardo, O., Sanz-Aparicio, J. y Díaz, P. in Industrial Enzymes: Structure, Function and Applications (2007) Chpt. 5, 65-82, Springer NL, Polaina, Julio; MacCabe, Andrew P. (Eds.), XII, 642 p., Hardcover. ISBN: 978-1-4020-5376-4 Role of the molecular conformation in the two- and three dimensional supramolecular structure of ten hydroxyl-N-alkylamides Foces-Foces, C., López-Rodríguez, M., Pérez, C., Martín, J.D. and Pérez-Hernández, N. Crystal Growth & Design (2007) 7, 905-911 Hydrogen bonding in 3-methylenecyclohexane-1,1-dicarboxylic acid C. Foces-Foces, M. L. Rodríguez and N. Pérez-Hernández Acta Crystallographica (2007) E63, o1308-o1310 2,6-Dimethyl-9-oxabicyclo[3.3.1]nonane-endo-2,endo-6-dio M. López-Rodríguez and C. Foces-Foces Acta Crystallographica (2007) E63, o3537-o3538 Solvent-free thermal and microwave-assisted [3 + 2] cycloadditions between stabilized azomethine ylides and nitrostyrenes. An experimental and theoretical study Arrieta A, Otaegui D, Zubia A, Cossío FP, Díaz-Ortiz A, de la Hoz A, Herrero MA, Prieto P, Foces-Foces C, Pizarro JL, and Arriortua MI The Journal of Organic Chemistry (2007) 72, 4313-4322 2006 A pharmacological map of the PI3-K family defines a role for p110α in insulin signaling Knight Z.A., González B., Feldman M.E., Zunder E.R., Goldenberg D.D., Williams O., Loewith R., Stokoe D., Balla A., Toth B., Balla T., Weiss W.A., Williams R.L. and Shokat K.M. Cell (2006) 125, 733-747 [PDF-copy] The role of electrostatic interactions in the antitumor activity of dimeric Rnases Notomista, E, Mancheño JM, Crescenzi, O, Di Donato A, Gavilanes, JG, D´Alessio, G FEBS J. (2006) 273, 3687-3697 A complementary microscopy analysis of Sticholysin II crystals on lipid films: Atomic Force and Transmission Electron characterizations Mancheño JM, Martín-Benito J, Gavilanes JG, Vázquez L Biophys. Chem. (2006) 119(3), 219-223 Chemical modification of the protein surfaces to improve their reversible enzyme immobilization on ionic exchangers Tamara Montes, Valeria Grazu, Fernando López-Gallego, Juan A. Hermoso, José M. Guisán and Roberto Fernández-Lafuente Biomacromolecules (2006). 7, 3052-3058 Cell signaling and function organized by PB1 domain interactions Moscat, J., Diaz-Meco, M.T., Albert, A. and Campuzano, S. Molecular Cell (2006) 23, 631-640 [PDF-copy] X-ray and Neutron Diffraction Approaches to the Structural Analysis of Protein-Lipids Interactions Juan A. Hermoso, José M. Mancheño and Eva Pebay-Peyroula in Protein-Lipid Interactions. New Approaches and Emerging Concepts (2006). Springer Verlag Berlin Heidelberg, pp.63-100 La Quimica de la vida a escala atomica Hermoso, J.A., Sanz-Aparicio, J.A. and Albert, A. Anales de Química (2006) 102(4), 15-22 [PDF-copy] Thermochemistry of 2,5-thiophenedicarboxylic acid Roux MV, Temprado M, Jiménez P, Foces-Foces C, Notario R, Verevkin SP, and Liebman JF The Journal of Physical Chemistry A (2006) 110, 12477-12483 Classification of Hydrogen-bond Motives in Crystals of N-H pyrazoles: a mixed empirical and theoretical approach Alkorta, I., Elguero, J., Foces-Foces, C. and Infantes, L. ARKIVOC(ii) (2006) 15-30 Síntesis and Structure of Hydroxyl Acids of General Structure 7,7-Alkenyl-5-hydroxymethyl-6-oxabicyclo[3.2.1octane-1-carboxylic Acid Pérez-Hernández, N., Febles, M., Pérez, C., Pérez, R., Rodríguez, M.L., Foces-Foces, C. and Martín, J.D. J.Org.Chem. (2006) 71, 1139-1151 Distinct Dynamic Behaviors of Water Molecules in Hydrated Pores Febles, M., Pérez-Hernández, N., Pérez, C., Pérez, R., Rodríguez, M.L., Foces-Foces, C. Roux, M.V., Morales, E.Q., Buntkowsky, G., Limbach, H.-H and Martín, J.D. J. Am.Chem.Soc. (2006) 128, 10008-10009 Microwave assisted synthesis and crystal structures of 2-imidazolines and imidazoles Hoz, A., Diaz-Ortiz, A., Mateo, M.C., Moral, M., Moreno, A., Elguero, J., Foces-Foces, Rodríguez, M.L. and Sánchez-Migallón, A. Tetrahedron (2006) 62, 5868- 5874 Supramolecular assembly of diethyl 1H-pyrazole-3,5-dicarboxylate 0.33-hydrate Foces-Foces, C., Rodríguez, M.L. and Elguero, J. Acta Cryst. (2006) E62, 03351-03353 Self-Associative Periodic Table of Elements by Neural Networks Fayos, J. in The Mathematics of the Periodic Table (2006) Rouvray, D.H. and King, R.B. ed. (Nova Science Publishers,Inc.) Chapter 6, pp 101-117 2005 Insights into pneumococcal pathogenesis from crystal structure of the modular teichoic acid phosphorylcholine esterase Pce Hermoso, J., Lagartera, L., Gonzalez, A., Stelter, M., Garcia, P., Martinez-Ripoll, M., Garcia, J.L. and Menendez, M. Nature Structural and Molecular Biology (2005) 12, 533-538 (doi:10.1038/nsmb940) C-terminal Tyrosine of Ferredoxin-NADP+ reductase in the hydride transfer processes with NAD(P)+/H J. Tejero, I. Pérez-Dorado, C. Maya, M. Martínez-Júlvez, J. Sanz-Aparicio, C. Gómez-Moreno, J.A. Hermoso and M. Medina Biochemistry (2005) 44, 13477-13490 Structure of the Functional Domain of phi 29 Replication Organizer: INSIGHTS INTO OLIGOMERIZATION AND DNA BINDING J.L. Asensio, A. Albert, D. Muñoz-Espín, C. González, J. Hermoso, L. Villar, J. Jiménez-Barbero, M. Salas, and W.J.J. Meijer Journal of Biological Chemistry (2005) 280, 20730-20739 Hydralysins: A new category of beta-pore-forming toxins in cnidaria. Characterization and preliminary structure-function analysis Sher DJ, Fishman Y, Zhang M, Lebendiker M, Gaathon A, Mancheño JM, Zlotkin E J. Biol. Chem. (2005) 280(24), 22847-22855 BzdR, a repressor that controls the anaerobic catabolism of benzoate in Azoarcus sp. CIB, is the first member of a new subfamily of transcriptional regulators Barragán MJ, Blázquez B, Zamarro MT, Mancheño JM, García JL, Díaz E, Carmona M J. Biol. Chem. (2005) 280(11), 10683-10694 The 1.49 A resolution crystal structure of PsbQ from photosystem II of Spinacia oleracea reveals a PPII structure in the N-terminal region Balsera. M., Arellano, J.B., Revuelta, J.L., de las Rivas, J. and Hermoso, J. Journal of Molecular Biology (2005). 350, 1051-1060 Crystallization and preliminary X-ray diffraction studies of the pneumococcal teichoic acid phosphorylcholine esterase Pce Lagartera, L., Gonzalez, A., Stelter, M., Garcia, P., Kahn, R., Menendez, M. and Hermoso, J. Acta Crystallographica (2005) F61, 221-224 (doi:10.1107/S1744309105001636) Conformational engineering of lipases via directed immobilization. Improving the resolution of chiral drugs J.M Palomo, G. Fernández-Lorente, C. Ortiz, R. L. Segura, C. Mateo, M. Fuentes, J. Hermoso, R. Fernández- Lafuente and J. M Guisán Medicinal Chemistry reviews-online (2005) 2(5), 369-378 [Web-access] Structural Analysis of the Laetiporus sulphureous Hemolytic Pore-forming Lectin in Complex with Sugars Mancheño, J.M., Tateno, H., Goldstein, I.J., Martínez-Ripoll, M. and Hermoso, J.A. Journal of Biological Chemistry (2005) 280, 17251-17259 Activation in the family of Candida rugosa isolipases by polyethylene glycol Otero, C., Fernández-Pérez, M., Hermoso, J.A. and Martínez-Ripoll, M. Journal of Molecular Catalysis B: Enzymatic (2005) 32, 225-229 Structural Analysis of Interactions for Complex Formation between Ferredoxin-NADP+ Reductase and Its Protein Partners Mayoral, T., Martinez-Julvez, M., Perez-Dorado, I., Sanz-Aparicio, J., Gomez-Moreno, C., Medina, M. and Hermoso, J.A. Proteins: Structure, Function, and Bioinformatics (2005) 59, 592-602 Pneumococcal phosphorylcholine esterase, Pce, contains a zinc binuclear center that is essential for substrate binding and catalysis L. Lagartera, A. González, J. A. Hermoso, J. L. Saíz, P. García, J. L. García and M. Menéndez Protein Science (2005) 14, 3013-3024 (doi:10.1110/ps.051575005) The Ferredoxin-NADP(H) Reductase from Rhodobacter capsulatus: Molecular Structure and Catalytic Mechanism I. Nogués, I. Pérez-Dorado, S. Frago, C. Bittel, S. G. Mayhew, C. Gómez-Moreno, J. A. Hermoso, M. Medina, N. Cortez, and N. Carrillo Biochemistry (2005) 44, 11730-11740 Probing the Determinants of Substrate Specificity of a Feruloyl Esterase, AnFaeA, from Aspergillus niger C.B. Faulds, R. Molina, R. Gonzalez, F. Husband, N. Juge, J. Sanz-Aparicio and J. A. Hermoso FEBS Journal (2005) 272, 4362-4371 The structure of the functional dimeric domain of phage f29 protein p16.7 reveals mechanisms for oligomerization and DNA binding Asensio, JL, Albert, A, Muñoz-Espín, D, González, C, Hermoso, JA, Villar, L, Jiménez-Barbero, J, Salas, M and Meijer, W Journal of Biological Chemistry (2005) 280, 20730-20739 Structural basis for membrane anchorage of viral phi 29 DNA during replication Albert A, Munoz-Espin D, Jimenez M, Asensio JL, Hermoso JA, Salas M, Meijer WJ. Journal of Biological Chemistry (2005) 280, 42486-42489 Sánchez-Barrena, M.J., Martínez-Ripoll, M., Zhu, J.K. and Albert, A. The structure of the Arabidopsis thaliana SOS3: molecular mechanism of sensing calcium for salt stress response Journal of Molecular Biology (2005) 345, 1253-1264 Rat liver betaine homocysteine S-methiltransferase equilibrium unfolding: Insights into intermediate structure through tryptophan substitutions Garrido, F., Gasset, M., Sanz-Aparicio, J., Alfonso, C. y Pajares, M.A. Biochemical Journal (2005) 391, 589-599 2004 ESCRT-II, an endosome-associated complex required for protein sorting: crystal structure and interactions with ESCRT-III and membranes. Teo H., Perisic O., González B. and Williams R.L. Developmental Cell (2004) 7, 559-569 The structure of human inositol 1,4,5-trisphosphate 3-kinase: The substrate binding reveals why it is not a phosphoinositide 3-kinase González B., Schell M.J., Letcher, A.J., Veprintsev, D.B., Irvine R. and Williams R.L. Molecular Cell (2004) 15,689-701 SOS3 (salt overlay sensitive 3) from Arabidopsis thaliana: expression, purification, crystallization and preliminary X-ray analysis Sánchez-Barrena, M.J., Martínez-Ripoll, M., Zhu, J.K. and Albert, A. Acta Crystallographica (2004) D60, 1272-1274 Structure of concanavalin A at pH 8: bound solvent and crystal contacts Lopez-Jaramillo FJ, Gonzalez-Ramirez LA, Albert A, Santoyo-Gonzalez F, Vargas-Berenguel A, Otalora F Acta Crystallographica (2004) D60, 1048-1046 Peptide AS-48: prototype of a new class of cyclic bacteriocins Maqueda M, Galvez A, Bueno MM, Sanchez-Barrena MJ, Gonzalez C, Albert A, Rico M, Valdivia E. Current Protein and Peptide Science (2004) 5, 399-416 Crystallization and preliminary crystallographic analysis of a novel haemolytic lectin from the mushroom Laetiporus sulphureus Mancheño, J.M., Hiroaki, T., Goldstein, I.J. and Hermoso, J.A. Acta Crystallographica (2004) D60, 1139-1141 Crystallization of a proteolyzed form of the horse pancreatic lipase-related protein 2: structural basis for the specific detergent requirement Mancheño, J.M., Jayne, S., Kerfelec, B., Chapus, C., Crenon, I. and Hermoso, J.A. Acta Crystallographica (2004) D60, 2107-2109 Crystallization and preliminary X-ray diffraction analysis of ferredoxin-NADP(H) reductase from Rhodobacter capsulatus Perez-Dorado. I., Bittel, C., Cortez, N. and Hermoso, J.A. Acta Crystallographica (2004) D60, 2332-2335 Stabilization of Penicillin G Acylase from Escherichia coli: Site-Directed Mutagenesis of the Protein Surface To Increase Multipoint Covalent Attachment Abian, O., Grazu, V., Hermoso, J.A., Gonzalez, R., Garcia, J.L., Fernandez-Lafuente, R. and Guisan, J.M. Applied and Environmental Microbiology (2004) 70, 1249-1251 Alegre-Cebollada, J, Lacadena, V, Oñaderra, M, Mancheño, JM, Gavilanes JG, Martínez del Pozo, A Phenotypic selection and characterization of randomly produced non-haemolytic mutants of the toxic sea anemone protein Sticholysin II FEBS Lett. (2004) 575, 14-18 The Crystal Structure of Feruloyl Esterase A from Aspergillus niger Suggests Evolutive Functional Convergence in Feruloyl Esterase Family Hermoso, J.A., Sanz-Aparicio, J., Molina, R., Juge, N., Gonzalez, R. and Faulds, C.B. Journal of Molecular Biology (2004) 338, 495-506 Functional characterization of the yeast Ppz1 phosphatase inhibitory subunit Hal3: a mutagenesis study Muñoz, I, Ruiz, A, Marquina, M, Barcelo, A, Albert, A, Ariño, J. Journal of Biological Chemistry (2004) 279, 42619-42627 2003 Estructura de proteínas por Cristalografía de rayos X Hermoso, J.A. y Martínez-Ripoll, M. in Estructura de proteínas (2003). Gómez-Moreno, C. and Sancho Sanz, J., coord., Ariel Ciencia, ISBN: 84-344-8061-6, pp. 253-285 Crystallization and preliminary X-ray diffraction studies of two different crystal forms of the lipase 2 isoform from the yeast Candida rugosa Mancheño, J.M., Pernas, M.A., Rua, M.L. and Hermoso, J.A. Acta Crystallographica (2003) D59, 499-501 Structural aspects on rat liver S-adenosylmethionine synthesis Pajares, M.A., Gasset, M., Sanz-Aparicio, J., Calvete, J.J. and Rodriguez Arrondo, J.L. Advances in Biological Chemistry (2003) 2, 31-41 Active-site-mutagenesis study of rat liver betaine-homocysteine S-methyltransferase González, B., Campillo, N., Garrido, F., Gasset, M., Sanz-Aparicio, J. and Pajares, M.A. Biochem. J. (2003) 370, 945-952 Involvement of the Pyrophosphate and the 2'-Phosphate Binding Regions of Ferredoxin-NADP+ Reductase in Coenzyme Specificity Tejero, J., Martinez-Julvez, M., Mayoral, T., Luquita, A., Sanz-Aparicio, J., Hermoso, J.A., Hurley, J.K., Tollin, G., Gómez-Moreno, C. and Medina, M. Journal of Biological Chemistry (2003) 278, 49203-49214 Structure of Bacteriocin AS-48: From Soluble State to Membrane Bound State Sánchez-Barrena, M.J., Martinez-Ripoll, M., Gálvez, A., Valdivia, E., Maqueda, M., Cruz, V. and Albert, A. Journal of Molecular Biology (2003) 334, 541-549 Structural Insights into the Lipase/esterase Behavior in the Candida rugosa Lipases Family: Crystal Structure of the Lipase 2 Isoenzyme at 1.97 A Resolution Mancheño, J.M., Pernas, M.A., Martínez, M.J., Ochoa, B., Rúa, M.L. and Hermoso, J.A. Journal of Molecular Biology (2003) 332, 1059-1069 Crystal Structures of Methionine Adenosyltransferase Complexed with Substrates and Products Reveal the Methionine-ATP Recognition and Give Insights into the Catalytic Mechanism González, B., Pajares, M.A., Hermoso, J.A., Guillerm, D., Guillerm, G. and Sanz-Aparicio, J. Journal of Molecular Biology (2003) 331, 407-416 Structural Basis for Selective Recognition of Pneumococcal Cell Wall by Modular Endolysin from Phage Cp-1 Hermoso, J.A., Monterroso, B., Albert, A., Galán, B., Ahrazem, O., García, P., Martínez-Ripoll, M., García, J.L., and Menéndez, M. Structure (2003) 11, 1239-1249 Crystal and Electron Microscopy Structures of Sticholysin II Actinoporin Reveal Insights into the Mechanism of Membrane Pore Formation. Mancheño, J.M., Martín-Benito, J., Martínez-Ripoll, M., Gavilanes, J.G. and Hermoso, J.A. Structure (2003) 11, 1319-1328 2002 Crystallization and preliminary X-ray study of recombinant betaine-homocysteine S-methyltransferase from rat liver González, B., Pajares, M.A., Too, H.-P., Garrido, F., Blundell, T.L. and Sanz-Aparicio, J. Acta Crystallographica (2002) D58, 1507-1510 Crystallization and preliminary X-ray diffraction studies of the complete modular endolysin from Cp-1, a phage infecting Streptococcus pneumoniae Monterroso, B., Albert, A., Martínez-Ripoll, M., García, P., García, J.L., Menéndez, M. and Hermoso, J.A. Acta Crystallographica (2002) D58, 1487-1489 Crystallization and preliminary X-ray diffraction studies of the water-soluble state of the pore-forming toxin sticholysin II from the sea anemone Stichodactyla helianthus Mancheño, J.M., Martínez-Ripoll, M., Gavilanes, J.G. and Hermoso, J.A. Acta Crystallographica (2002) D58, 1229-1231 Activation of Horse PLRP2 by Bile Salts Does Not Require Colipase Jayne, S., Kerfelec, B., Foglizzo, E., Granon, S., Hermoso, J.A., Chapus, C. and Crenon, I. Biochemistry (2002) 41, 8422-8428 Structural basis for the kinetics of Candida rugosa Lip1 and Lip3 isoenzymes Pernas, M., López, C., Prada, A., Hermoso, J. and Rúa, M.L. Colloids and Surfaces B: Biointerfaces (2002) 26, 67-74 Probing the role of glutamic acid 139 of Anabaena ferredoxin-NADP+ reductase in the interaction with substrates Faro, M., Frago, S., Mayoral, T., Hermoso, J.A., Sanz-Aparicio, J., Gómez-Moreno, C. and Medina, M. European Journal of Biochemistry (2002) 269, 4938-4947 TrwD, the Hexameric Traffic ATPase Encoded by Plasmid R388, Induces Membrane Destabilization and Hemifusion of Lipid Vesicles Machón, C., Rivas, S., Albert, A., Goñi, F.M. and de la Cruz, F. Journal of Bacteriology (2002) 184, 1661-1668 Mechanism of Coenzyme Recognition and Binding Revealed by Crystal Structure Analysis of Ferredoxin-NADP+ Reductase Complexed with NADP+ Hermoso, J.A., Mayoral, T., Faro, M., Gómez-Moreno, C., Sanz-Aparicio, J. and Medina, M. Journal of Molecular Biology (2002) 319, 1193-1142 Estructural Enzymology of Li+-sensitive/Mg2+-dependent Phosphatases Patel, S., Martinez-Ripoll, M., Blundell, T.L. and Albert, A. Journal of Molecular Biology (2002) 320, 1087-1094 2001 Probing the Determinants of Coenzyme Specificity in Ferredoxin-NADP+ Reductase by Site-directed Mutagenesis Medina, M., Luquita, A., Tejero, J., Hermoso, J., Mayoral, T., Sanz-Aparicio, J., Grever, K. and Gómez-Moreno, C. Journal of Biological Chemistry (2001) 276, 11902-11912 Role of a Cluster of Hydrophobic Residues Near the FAD Cofactor in Anabaena PCC 7119 Ferredoxin-NADP+ Reductase for Optimal Complex Formation and Electron Transfer to Ferredoxin Martínez-Júlvez, M., Nogués, I., Faro, M., Hurley, J.K., Brodie, T.B., Mayoral, T., Sanz-Aparicio, J., Hermoso, J., Stankovich, M.T., Medina, M., Tollin, G. and Gómez-Moreno, C. Journal of Biological Chemistry (2001) 276, 27498-27510 Regioselective resolution of 1,n-diols catalysed by lipases: a rational explanation of the eznymatic selectivity Borreguero, I., Sánchez-Montero, J.M., Sinisterra, J.V., Rumbero, A., Hermoso, J. and Alcántara, A.R. Journal of Molecular Catalysis B: Enzymatic (2001) 11, 1013-1024 Influence of the conformational flexibility on the kinetics and dimerisation process of two Candida rugosa lipase isoenzymes Pernas, M.A., López, C., Rúa, M.L. and Hermoso, J. FEBS Letters (2001), 501, 87-91 2000 The X-ray Structure of the FMN-binding Protein AtHal3 Provides the Structural Basis for the Activity of a Regulatory Subunit Involved in Signal Transduction Albert, A., Martinez-Ripoll, M., Espinosa-Ruiz, A., Yenush, L., Culiañez-Macia, F.A. and Serrano, R. Structure (2000) 8, 961-969 Critical Role of Micelles in Pancreatic Lipase Activation Revealed by Small Angle Neutron Scattering Pignol, D., Ayvazian, L., Kerfelec, B., Timmins, P., Crenon, I., Hermoso, J., Fontecilla-Camps, J.C. and Chapus, C. Journal of Biological Chemistry (2000) 275, 4220-4224 Direct Evolution of β-Glucosidase A from Paenibecillus polymyxa to Thermal Resistance Gonzalez-Blasco, G., Sanz-Aparicio, J., Gonzalez, B., Hermoso, J.A. and Polaina, J. Journal of Biological Chemistry (2000) 275, 13708-13712 X-ray Structure of Yeast Hal2p, a Major Target of Lithium and Sodium Toxicity, and Identification of Framework Interactions Determining Cation Sensitivity Albert, A., Yenush, L., Gil-Mascarell, M.R., Rodriguez, P.L., Patel, S., Martinez-Ripoll, M., Blundell, T.L. and Serrano, R. Journal of Molecular Biology (2000) 295, 927-938 The Crystal Structure of Tetrameric Methionine Adenosyltransferase from Rat Liver Reveals the Methionine-binding site González, B., Pajares, M.A., Hermoso, J.A., Alvarez, L., Garrido, F., Sufrin, J.R. and Sanz-Aparicio, J. Journal of Molecular Biology (2000) 300, 363-375 Structural Basis of the Catalytic Role of Glu301 in Anabaena PCC 7119 Ferredoxin-NADP+ Reductase Revealed by X-Ray Crystallography Mayoral, T., Medina, M., Sanz-Aparicio, J., Gomez-Moreno, C. and Hermoso, J. Proteins: Structure, Function, and Genetics (2000) 38, 60-69 Biochemical and Physical Characterization of Parvovirus Minute Virus of Mice Virus-like Particles Hernando, E., Llamas-Saiz, A.L., Foces-Foces, C., McKenna, R., Portman, I., Agbandje-McKenna, M. and Almendral, J.M. Virology (2000) 267, 299-309 1999 Making the most of commercial sparse-matrix protein crystallization screening kits Albert, A. and Martinez-Ripoll, M. Journal of Applied Crystallography (1999) 32, 336-338 Molecular study of the rat liver NADH: Cytochrome c oxidoreductase complex during development and ageing Torres-Mendoza, C., Albert, A. and Cruz Arriaga, M. Molecular and Cellular Biochemistry (1999) 195, 133-142 A six-stranded double-psi beta barrel is shared by several protein superfamilies Castillo, R.M., Mizuguchi, K., Dhanaraj, V., Albert, A., Blundell, T.L. and Murzin, A.G. Structure (1999) 7, 227-236 Acyclic Phenylalkadienols as Substrates for the Study of Enzyme Recognition. Regioselective Acylation by Porcine Pancreatic Lipase: A Structural Hypothesis for the Enzymatic Activity Borreguero, I., Sinisterra, J.V., Rumbero, A., Hermoso, J., Martinez-Ripoll, M. and Alcantara, A.R. Tetrahedron (1999), 55, 14961-14974 1998 Pancreatic Lipase-Related Protein Type 1: A Double Mutation Restores a Significant Lipase Activity Crenon, I., Jayne, S., Kerfelec, B., Hermoso, J., Pignol, D. and Chapus, C. Biochemical and Biophysical Research Communications (1998) 246, 513-517 Role of Arg100 and Arg264 from Anabaena PCC 7119 Ferredoxin-NADP+ Reductase for Optimal NADP+ Binding and Electron Transfer Martinez-Julvez, M., Hermoso, J., Hurley, J.K., Mayoral, T., Sanz-Aparicio, J., Tollin, G., Gomez-Moreno, C. and Medina, M. Biochemistry (1998) 37, 17680-17691 The lipase/colipase complex is activated by a micelle: neutron crystallographic evidence Pignol, D., Hermoso, J., Kerfelec, B., Crenon, I., Chapus, C. and Fontecilla-Camps, J.C. Chemistry and Physics of Lipids (1998) 93, 123-129 Ion Pairing between Lipase and Colipase Plays a Critical Role in Catalysis Ayvazian, L., Crenon, I., Hermoso, J., Pignol, D., Chapus, C. and Kerfelec, B. Journal of Biological Chemistry (1998) 273, 33604-33609 Crystal Structure of beta-Glucosidase A from Bacillus polymyxa: Insights into the Catalytic Activity in Family 1 Glycosyl Hydrolases Sanz-Aparicio, J., Hermoso, J.A., Martinez-Ripoll, M., Lequerica, J.L. and Polaina, J. Journal of Molecular Biology (1998) 275, 491-502 Crystal structure of aspartate decarboxylase at 2.2 A resolution provides evidence for an ester in protein self-processing Albert, A., Dhanaraj, V., Genshel, U., Khan, G., Ramjee, M.K., Pulido, R., Sibanda, B.L., Delft, F., Witty, M., Blundell, T.L., Smith, A.G. and Abell, C. Nature Structural Biology (1998) 5, 289-293 Pancreatic Lipase related proteins type I: Specialized lipase or an inactive enzyme? Crenon, I., Flogizzo, E., Kerfelec, B., Virine, A., Pignol, D., Hermoso Dominguez, J., Bonicel, J. and Chapus, C. Protein Engineering (1998) 112, 135-142 Structural Basis of Increased Resistance to Thermal Denaturation Induced by Single Amino Acid Substitution in the Sequence of β-Glucosidase A From Bacillus polymyxa Sanz-Aparicio, J., Hermoso, J.A., Martinez-Ripoll, M., Gonzalez, B., Lopez-Camacho, C. and Polaina, J. Proteins: Structure, Function, and Genetics (1998) 33, 567-576 Functional implications of the structure of the murine parvovirus, minute virus of mice Agbandje-McKenna, M., Llamas-Saiz, A.L., Wang, F., Tattersall, P. and Rossmann, M.G. Structure (1998) 6, 1369-1381 1997 Structure Determination of Minute Virus of Mice Llamas-Saiz, A.L., Agbandje-McKenna, M., Wikoff, W.R., Bratton, J., Tattersall, P. and Rossmann, G. Acta Crystallographica (1997) D53, 93-102 Purification, Crystallization and Preliminary X-ray Diffraction Studies of C-Phycocyanin and Allophycocyanin from Spirulina platensis Moreno, A., Bermejo, R., Talavera, E., Alvarez-Pez, J.M., Sanz-Aparicio, J. and Romero, A. Acta Crystallographica (1997) D53, 321-326 Aspartic proteinases: From the first X-ray photos of pepsin crystals to hundreds of 3-D structures Blundell, T.L., Guruprasad, K., Albert, A., Williams, M., Sibanda, B.L. and Dhanaraj, V. Current Science (1997) 72, 483-489 Neutron Crystallographic Evidence of Lipase-Colipase Complex Activation by a Micelle Hermoso, J., Pignol, D., Roth, M., Chapus, C. and Fontecilla-Camps, J.C. EMBO Journal (1997) 16, 5531-5536 The Crystal Structure of Canavalia brasiliensis Lectin Suggest a Correlation Between its Quaternary Conformation and its Distinct Biological Properties from Concanavalin A Sanz-Aparicio, J., Hermoso, J.A., Grangeiro, T.B., Calvete, J.J. and Cavada, B.S. FEBS Letters (1997) 405, 114-118 1996 Lipase Activation by Nonionic Detergents. The Crystal Structure of the Porcine Lipase-Colipase-Tetraethylene Glycol Monooctyl Ether Complex Hermoso, J.A., Pignol, D., Kerfelec, B., Crenon, I., Chapus, C. and Fontecilla-Camps, J.C. Journal of Biological Chemistry (1996) 271, 18007-18016 Protein engineering loops in aspartic proteinases: site-directed mutagenesis, biochemical characterization and X-ray analysis of chymosin with a replaced loop from rhizopuspepsin Nugent, P.G., Albert, A., Orprayoon, P., Pitts, J.E., Blundell, T.L. and Dhanaraj, V. Protein Engineering (1996) 9, 885-893 Structural Analysis of a Mutation in Canine Parvovirus Which Controls Antigenicity and Host Range Llamas-Saiz, A.L., Agbandje-Mckenna, M., Parker, J.S.L., Wahid, A.T.M., Parrish, C.R. and Rossmann, M.G. Virology (1996) 225, 65-71 1993-1995 Macromoléculas biológicas: Estructura tridimensional Martínez-Ripoll, M., Romero, A. and Sanz-Aparicio, J. Investigación y Ciencia (1995) 222, 83-84 Crystallizacion and Preliminary X-ray Diffraction Analysis of a Type I β-Glucosidase Encoded by the bglA Gene of Bacillus polymyxa Sanz-Aparicio, J., Romero, A., Martinez-Ripoll, M., Madarro, A., Flors, A. and Polaina, J. Journal of Molecular Biology (1994) 240, 267-270 The Use of Protein Homologues in the Rotation Function Aguilar, C.F., Newman, M.P., Sanz-Aparicio, J., Cooper, J.B., Tickle, I.J. and Blundell, T.L. Acta Crystallographica (1993) A49, 306-315 |
| Facilities | |
The laboratory is very well equipped with a wide range of crystallographic and computing equipment, wet laboratories and dedicated facilities for protein production, protein characterisation, protein crystallization, X-ray diffraction and computing. A summary of all these facilities can be obtained through this link. Moreover (although only written in Spanish) some protocols for the labs of protein production and molecular biology can be obtained through this link. |
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| Protein production and Molecular Biology Laboratories |
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 Laboratory for protein production |
 Molecular Biology Lab
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| Crystallization Laboratory |
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 Crystallization Lab |
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 Crystallization platform |
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 Crystallization robot |
 Crystal-Farm
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| X-ray Diffraction Laboratory |
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 Kappa goniometer with CCD and rotating anode with mirrors |
 Imaging plate mounted on a rotating anode with mirrors |
 Kappa goniometer with Axiom detector and X-ray micro-source |
 Kappa goniometer with Axiom detector and X-ray micro-source |
| Computing Laboratory |
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 Graphic computers with stereoview |
 Graphic computers with stereoview |
| In
addition, there are several low and high pressure
chromatography systems and a large number of different chromatographic
matrices for optimal protein purification. Analytical techniques,
including electrophoresis are also available in the lab, which combined
with access to mass spectrometry and analytical ultracentrifugation are
used to assess the quality of the proteins produced. In addition,
through our links with remaining groups of the Institute, we
have access
to isothermal titration calorimetry, Nuclear
Magnetic Resonance, analytical ultracentrifugation and MALDI-TOF mass
spectrometry. Biochemical studies and assays can be performed using a number of techniques, such as UV-VIS and CD-spectroscopies, microcalorimetry and fluorimetry. The ultimate goal is to provide a regular supply of crystals for X-ray crystallographic structure determination. Success in this aim is aided by the presence of a number of temperature-controlled incubators, high-throughput facilities and dedicated crystallisation rooms. |
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