Structural study of a single‐point mutant of Sulfolobus solfataricus alcohol dehydrogenase with enhanced activity

FEBS Letters

Sica, Filomena¹  Raia, Carlo Antonio²  Esposito, Luciana¹  Rossi, Mosè²  Bruno, Ilaria³  Mazzarella, Lelio¹  Zagari, Adriana¹  Giordano, Antonietta² 

[1] Istituto di Biostrutture e Bioimmagini, CNR, via Mezzocannone 6-8, I-80134 Napoli, Italy;Istituto di Biochimica delle Proteine, CNR, via Marconi 10, I-80125 Napoli, Italy;Dipartimento di Chimica, Università degli Studi di Napoli ‘Federico II’, via Cinthia, I-80126 Napoli, Italy

关键词: Crystal structure;    Alcohol dehydrogenase;    Single mutation;    Enzyme activation;    Coenzyme binding;    ADH;    alcohol dehydrogenase;    SsADH;    Sulfolobus solfataricus ADH;    HLADH;    horse liver ADH;    wt;    wild-type;    NCS;    non-crystallographic symmetry;   

DOI  :  10.1016/S0014-5793(03)00173-X

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Alcohol dehydrogenase from Sulfolobus solfataricus (SsADH) is the only enzyme from Archaea among the structurally studied members of the medium-chain ADH family described so far. Here, we present the three-dimensional structure of the apo form of the mutant N249Y which exhibits increased catalytic activity when compared to the wild-type enzyme. The substitution, located in the coenzyme binding domain, decreases the affinity for NAD(H) cofactor. The rearrangement of segments 248–250 and 270–275, induced by the mutation, suggests an explanation for the lower coenzyme affinity. This study also highlights the role in SsADH catalysis of the flexible loops located at the interface between the catalytic and the coenzyme domains.

【 授权许可】

Unknown