Effect of protein disulfide isomerase on the rate‐determining steps of the folding of bovine pancreatic ribonuclease A

FEBS Letters

Shin, Hang-Cheol¹  Scheraga, Harold A¹  Song, Myeong-Cheol¹ 

[1] Baker Laboratory of Chemistry and Chemical Biology, Cornell University, Ithaca, NY 14853-1301, USA

关键词: Protein disulfide isomerase;    Ribonuclease A;    Protein folding;    Rate-determining steps;    Intermediates;    RNase A;    bovine pancreatic ribonuclease A;    PDI;    protein disulfide isomerase;    DTTox;    oxidized dithiothreitol;    DTTred;    reduced dithiothreitol;    AEMTS;    2-aminoethyl methanethiosulfonate;    des-[ ];    three-disulfide intermediate having three native disulfide pairings but lacking a disulfide bond specified within the brackets;   

DOI  :  10.1016/S0014-5793(02)02825-9

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

The effects of protein disulfide isomerase (PDI) on the four structured des species that accumulate in the rate-determining steps of ribonuclease A folding were investigated at pH 8.0 and 15°C. The results indicate that PDI catalyzes the conversion of the kinetically trapped intermediates, des-[26–84] and des-[58–110], by reshuffling them into the on-pathway intermediate, des-[40–95], and the formation of native protein, by acting as both a chaperone and an oxidase on this on-pathway intermediate. These results provide the first strong evidence for the mechanism of PDI in the rate-determining steps of the oxidative folding pathways of ribonuclease A. Our approach, using PDI and blocked PDI, combined with the fast-blocking 2-aminoethyl methanethiosulfonate method, may be generally applicable to the clarification of the effect of PDI on folding intermediates.

【 授权许可】

Unknown