Catalytic cysteine residues of ER‐60 protease

FEBS Letters

Moriyama, Tatsuya¹  Kito, Makoto¹  Okudo, Hirokazu¹  Urade, Reiko¹ 

[1] Research Institute for Food Science, Kyoto University, Uji, Kyoto 611-0011, Japan

关键词: Endoplasmic reticulum;    Cysteine protease;    Catalytic dyad;    Site-directed mutagenesis;    ER-60;    ER-60 protease;    ER;    endoplasmic reticulum;    pCMB;    p-chloromercuribenzoate;    DFP;    diisopropyl fluorophosphate;    PDI;    protein disulfide isomerase;    PCR;    polymerase chain reaction;    IPTG;    isopropyl-1-thio-β-D-galactopyranoside;    PAGE;    polyacrylamide gel electrophoresis;    CBB;    Coomassie brilliant blue R-250;    bis-Tris;    bis-(2-hydroxyethyl)iminotris (hydroxymethyl) methane;   

DOI  :  10.1016/S0014-5793(99)01721-4

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

ER-60 protease contains two CGHC motifs that appear to include an active site cysteine residue(s). Its proteolytic activity was lost with a double mutation of the C-terminal cysteines of the two motifs to alanine, but not with a single mutation of the C-terminal cysteine of either of the motifs to alanine. This suggests that these C-terminal cysteines independently constitute the catalytic active site. A mutation of both histidine residues in the two CGHC motifs to serine did not abolish the proteolytic activity, suggesting these histidine residues in the CGHC motifs do not constitute the catalytic dyad of ER-60 protease.

【 授权许可】

Unknown