title: Crystallization and preliminary X-ray analysis of NADP(H)-dependent alcohol dehydrogenases from Saccharomyces cerevisiae and Rana perezi.
abstract: 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 A resolution and belonged to the trigonal space group P3(1)21 (or to its enantiomorph P3(2)21), with unit-cell parameters a = b = 102.2, c = 149.7 A, gamma = 120 degrees. 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 A, respectively, using synchrotron radiation. These crystals were isomorphous, space group C2, with approximate unit-cell parameters a = 122, b = 79, c = 91 A, beta = 113 degrees and contain one dimer per asymmetric unit, with a Volume solvent content of about 50%.
authors.affiliation: IBMB-CSIC, Jordi Girona 18-26, E-08034 Barcelona, Spain.
authors.forename: Eva
authors.initials: E
authors.lastname: Valencia
authors.forename: Albert
authors.initials: A
authors.lastname: Rosell
authors.forename: Carolina
authors.initials: C
authors.lastname: Larroy
authors.forename: Jaume
authors.initials: J
authors.lastname: Farrés
authors.forename: Josep A
authors.initials: JA
authors.lastname: Biosca
authors.forename: Ignacio
authors.initials: I
authors.lastname: Fita
authors.forename: Xavier
authors.initials: X
authors.lastname: Parés
authors.forename: Wendy F
authors.initials: WF
authors.lastname: Ochoa
mesh_terms: D000426:Alcohol Dehydrogenase / Q000737:chemistry* / Q000378:metabolism; D000818:Animals; D003460:Crystallization / Q000379:methods; D018360:Crystallography, X-Ray; D008958:Models, Molecular; D009249:NADP / Q000378:metabolism*; D021122:Protein Subunits / Q000737:chemistry; D011898:Ranidae / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D017356:Synchrotrons
chemical_list: D021122:Protein Subunits; D009249:NADP; D000426:Alcohol Dehydrogenase