Evidence for an active‐center cysteine in the SH‐proteinase α‐clostripain through use of N‐tosyl‐L‐lysine chloromethyl ketone

FEBS Letters

Gilles, A.-M.¹  Keil, B.¹ 

[1] Unité de Chimie des Protéines, Institut Pasteur, 28, rue du Docteur Roux, 75724 Paris Cédex 15, France

关键词: α-Clostripain;    Cysteine proteinase;    Active site;    TLCK;    N-α-p-tosyl-L-lysine chloromethyl ketone;    TPCK;    N-α--p-tosyl-L-phenylalanine chloromethyl ketone;    PMSF;    phenylmethylsulfonyl fluoride;    BAEE;    α-N-benzyl-L-arginine ethyl ester;    DTT;    dithiothreitol;    DTNB;    5;    5'-dithiobis(2-nitrobenzoic acid);    p-NO2-ZACK;    N-α-p-nitrobenzyloxycarbonyl-L-arginine chloromethyl ketone;   

DOI  :  10.1016/0014-5793(84)81017-0

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

The rapid reaction of α-clostripain with tosyl-L-lysine chloromethyl ketone results in a complete loss of activity and in the disappearance of one titratable SH group whereas the number of histidine residues is not affected. Tosyl-L-phenylalanine chloromethyl ketone and phenylmethylsulfonyl fluoride have no effect on the catalytic activity. From the molar ratio and under the assumption of 1:1 molar interaction, the fully active enzyme has a specific activity of 650–700 [twice the value proposed by Porter et al. (J. Biol. Chem. 246 (1971) 7675-7682)]. Partial oxidation makes it experimentally impossible to attain this maximal value.

【 授权许可】

Unknown