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

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• 作者：Shin ; Hang-Cheol ; Song ; Myeong-Cheol ; Scheraga ; Harold A.
• 关键词：Protein disulfide isomerase ; Ribonuclease A ; Protein folding ; Rate-determining steps ; Intermediates ; RNase A ; bovine pancreatic ribonuclease A ; PDI ; protein disulfide isomerase ; DTT^ox ; oxidized dithiothreitol ; DTT^red ; reduced dithiot
• 刊名：FEBS Letters
• 出版年：2002
• 出版时间：June 19, 2002
• 年：2002
• 卷：521
• 期：1-3
• 页码：77-80
• 全文大小：137 K

文摘

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.