Crystallization and preliminary X-ray analysis of NADP(H)-dependent alcohol dehydrogenases from Saccharomyces cerevisiae and Rana perezi
Valencia, Eva (Institut de Biologia Molecular de Barcelona)
Rosell i Febres, Albert (Universitat Autònoma de Barcelona. Departament de Bioquímica i de Biologia Molecular)
Larroy Hernando, Carolina (Universitat Autònoma de Barcelona. Departament de Bioquímica i de Biologia Molecular)
Farrés, Jaume (Universitat Autònoma de Barcelona. Departament de Bioquímica i de Biologia Molecular)
Biosca Vaqué, José Antonio (Universitat Autònoma de Barcelona. Departament de Bioquímica i de Biologia Molecular)
Fita Rodríguez, Ignasi (Institut de Biologia Molecular de Barcelona)
Parés i Casasampera, Xavier (Universitat Autònoma de Barcelona. Departament de Bioquímica i de Biologia Molecular)
Ochoa, Wendy F. (Institut de Biologia Molecular de Barcelona)

Date:	2003
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 Å 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%.
Rights:	Tots els drets reservats.
Language:	Anglès
Document:	Article ; Versió publicada
Subject:	Cristal·lografia ; Crystallography ; NADP (H)-dependent alcohol dehydrogenases
Published in:	Acta crystallographica. Section D, Biological crystallography, Vol. 59, Issue 2 (February 2003) , p. 334-337, ISSN 1399-0047

DOI: 10.1107/S090744490201661X

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