Drosophila alcohol dehydrogenase: evaluation of Ser139 site‐directed mutants

FEBS Letters

Atrian, Sı́lvia¹  Cols, Neus¹  Ladenstein, Rudolf²  Benach, Jordi²  Gonzàlez-Duarte, Roser¹ 

[1] Department de Genètica, Facultat de Biologia, Universitat de Barcelona, Av. Diagonal 645, E-08028 Barcelona, Spain;Karolinska Institutet, NOVUM, Center of Structural Biochemistry. S-141 57 Huddinge, Sweden

关键词: Site-directed-mutagenesis;    Alcohol dehydrogenase;    Short-chain dehydrogenase/reductase;    Catalytic triad;    Drosophila;   

DOI  :  10.1016/S0014-5793(97)00894-6

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Drosophila alcohol dehydrogenase (DADH) belongs to the large and highly heterogeneous (15–30% residue identity) short-chain dehydrogenase/reductase family (SDR). It is the only reported member that oxidizes mainly ethanol and 2-propanol among other alcohols. To confirm the role of Ser¹³⁹ we constructed two site-directed mutants, Ser¹³⁹Ala and Ser¹³⁹Cys, which show no enzymatic activity. Molecular replacement and data from crystallographically refined 3D structures confirm the position of Ser¹³⁹, whose hydroxyl group faces the cleft of the presumed catalytic pocket, very close to Tyr¹⁵² and Lys¹⁵⁶. Thus, consistent with the constitution of the catalytic triad of other SDR, our results suggest that Ser¹³⁹ of DADH is directly involved in the catalytic reaction.

【 授权许可】

Unknown