Crystallization and preliminary X-ray analysis of NADP(H)-dependent alcohol dehydrogenases from Saccharomyces cerevisiae and Rana perezi.
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%.
Valencia
Eva
E
IBMB-CSIC, Jordi Girona 18-26, E-08034 Barcelona, Spain.
Rosell
Albert
A
Larroy
Carolina
C
Farrés
Jaume
J
Biosca
Josep A
JA
Fita
Ignacio
I
Parés
Xavier
X
Ochoa
Wendy F
WF
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
D021122:Protein Subunits; D009249:NADP; D000426:Alcohol Dehydrogenase