Introduction of a free cysteinyl residue at position 68 in the subtilisin Savinase, based on homology with proteinase K

FEBS Letters

Breddam, K.¹  Branner, S.²  Bech, L.M.¹  Hastrup, S.² 

[1] Carlsberg Laboratory, Department of Chemistry, Gamle Carlsbergvef 10, DK-2500 Valby, Denmark;Novo Nordisk A/S Novo Alle, DK-2880 Bagsværd, Denmark

关键词: Subtilisin;    Cystein residue;    Mercurial;    Savinase;    subtilisin 309 from Bacillus lentus;    MES;    2-[N-morpholino]ethane sulfonic acid;    V68C Savinase;    Savinase with a cysteinyl residue at position 68;    Ph-Hg-Cl;    phenyl mercuric chloride;    HEPES;    N-[2-hydroxyethyl]piperazine-N′-[2-ethanesulfonic acid];    Suc;    succinyl;    pNA;    p-nitroanilid;    BICINE;    N;    N-bis[2-hydroxyethyl]-glycine;    SDS;    sodium dodecylsulfate;    KNPU;    kilo novo protease units;   

DOI  :  10.1016/0014-5793(92)80351-G

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Two subfamilies of the subtilisins, distinguished by the presence or absence of a free cysteinyl residue near the essential histidyl residue of the catalytic triad, are known. In order to evaluate the significance of the presence of this -SH group a cysteinyl residue has been introduced by site-directed mutagenesis into the cysteine-free subtilisin-like enzyme from Bacillus lentus, i.e. Savinase. The free cysteine affects the enzyme activity only slightly but renders it sensitive to mercurials presumably due to an indirect effect. The results indicate that the -SH group is not involved in catalysis.

【 授权许可】

Unknown