Intra-A chain disulphide bond forms first during insulin precursor folding.

Y Yuan; Z H Wang; J G Tang

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PubMed · 10493922

Intra-A chain disulphide bond forms first during insulin precursor folding.

Y Yuan·Z H Wang·J G Tang

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Abstract

In this study, we investigated the folding pathway of insulin precursor and compared it with that of insulin-like growth factor I (IGF-I). The intra-A chain disulphide bond was found to form early in insulin precursor folding, whereas the corresponding disulphide bond in IGF-I formed late. Intra-A chain disulphide-bond deleted [A6, A11-Ser] proteins, including proinsulin, insulin, and A chain, were employed for this investigation. Under the same conditions the recombination yield of insulin from S-sulphonates of native A and B chains was 22%, while the yield of [A6, A11-Ser] insulin from S-sulphonates of [A6, A11-Ser] A chain and native B chains was only approx. 7%. This indicated that the intra-A chain disulphide bond may serve to stabilize the A chain folding intermediate so as to facilitate the correct recognition and pairing with the B chain. Time courses of oxidation of reduced insulin A chains, reduced A and B chains, and reduced proinsulins showed that the intra-A chain disulphide bond formed first during insulin precursor folding. The formation of intra-A chain disulphide bond further accelerated the formation of the other two inter-chain disulphide bonds. The time course of helix structure formation of insulin A chains also indicated that the intra-A chain disulphide bond formed first, and could stabilize partially folded A chain helix structure. The rate of intra-A chain disulphide bond formation was almost the same as that for both helix structure formation and insulin molecule formation, indicating that the formation of the intra-A chain disulphide bond was the rate limiting step for the folding of insulin precursor.

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Circular Dichroism Disulfides Insulin Insulin-Like Growth Factor I Oxidation-Reduction Protein Folding Protein Precursors Protein Structure, Secondary Journal Article Research Support, Non-U.S. Gov't

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Y Yuan, Z H Wang, J G Tang. 1999-10-01. Intra-A chain disulphide bond forms first during insulin precursor folding.. https://pubmed.ncbi.nlm.nih.gov/10493922/

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