Protein engineering of Drosophila alcohol dehydrogenase The hydroxyl group of Tyr152 is involved in the active site of the enzyme

FEBS Letters

Gonzålez-Duarte, Ricard Albalat¹  Atrian, Silvia¹ 

[1] Department de Gen≐tica, Facultat de Biologia, Universitat de Barcelona, Av. Diagonal 645, E 08028 Barcelona, Spain

关键词: Site-directed-mutagenesis;    Drosophila;    Alcohol dehydrogenase;    Reactive residue;   

DOI  :  10.1016/0014-5793(92)81282-Q

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Drosophila alcohol dehydrogenase is the most studied member of the family of short-chain alcohol dehydrogenases, although its tridimensional structure still remains unknown. We have engineered a Drosophila alcohol dehydrogenase in which tyrosine-152, an invariant residue in all members of the family, has been substituted by phenylalanine. The mutated gene has been expressed in yeast and pure mutant enzyme has ban prepared by a one-step FPLC chromatographic procedure, Drosophila alcohol dehydrogenase-phenylalanine-152 shows no enzymatic activity. This result suggests not only that tyrosine-152 could constitute an essential building block of the active site but also that its hydroxyl group is directly involved in the redox reaction catalyzed by the enzyme.

【 授权许可】

Unknown