Crystallization and preliminary X-ray analysis of NADP(H)-dependent alcohol dehydrogenases from Saccharomyces cerevisiae and Rana perezi
| Autor: | Jaume Farrés, Wendy F. Ochoa, Albert Rosell, Ignacio Fita, Carolina Larroy, Josep A. Biosca, Xavier Parés, Eva Valencia |
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| Rok vydání: | 2021 |
| Předmět: |
Models
Molecular Ammonium sulfate Ranidae Dimer Saccharomyces cerevisiae Cristal·lografia Dehydrogenase Reductase Crystallography X-Ray law.invention chemistry.chemical_compound Structural Biology law Animals Crystallization Crystallography biology Alcohol Dehydrogenase General Medicine Lithium sulfate biology.organism_classification Solvent Protein Subunits chemistry NADP (H)-dependent alcohol dehydrogenases NADP Synchrotrons |
| Zdroj: | Recercat: Dipósit de la Recerca de Catalunya Varias* (Consorci de Biblioteques Universitáries de Catalunya, Centre de Serveis Científics i Acadèmics de Catalunya) Digital.CSIC. Repositorio Institucional del CSIC instname Dipòsit Digital de Documents de la UAB Universitat Autònoma de Barcelona Recercat. Dipósit de la Recerca de Catalunya |
| Popis: | Different crystal forms diffracting to high resolution have been obtained for two NADP(H)-dependent alcohol dehydrogenases, members of the medium-chain dehydrogenase/reductase superfamily: ScADHVI from Saccharomyces cerevisiae and ADH8 from Rana perezi. ScADHVI is a broad-specificity enzyme, with a sequence identity lower than 25% with respect to all other ADHs of known structure. The best crystals of ScADHVI diffracted beyond 2.8 resolution and belonged to the trigonal space group P3121 (or to its enantiomorph P3221), with unit-cell parameters a = b = 102.2, c = 149.7 Å, γ = 120°. These crystals were produced by the hanging-drop vapour-diffusion method using ammonium sulfate as precipitant. Packing considerations together with the self-rotation function and the native Patterson map seem to indicate the presence of only one subunit per asymmetric unit, with a volume solvent content of about 80%. ADH8 from R. perezi is the only NADP(H)-dependent ADH from vertebrates characterized to date. Crystals of ADH8 obtained both in the absence and in the presence of NADP+ using polyethylene glycol and lithium sulfate as precipitants diffracted to 2.2 and 1.8 Å, respectively, using synchrotron radiation. These crystals were isomorphous, space group C2, with approximate unit-cell parameters a = 122, b = 79, c = 91 Å, β = 113° and contain one dimer per asymmetric unit, with a volume solvent content of about 50%. This work was supported by grants PB98-0855 to XP, BMC2000-0132 to JAB and BIO099-0865 to IF |
| Databáze: | OpenAIRE |
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