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. 1990 Jun 15;268(3):803–806. doi: 10.1042/bj2680803

Protein folding/refolding analysis by mass spectrometry. Scrambling of disulphide bridges in insulin.

H R Morris 1, P Pucci 1, M Panico 1, G Marino 1
PMCID: PMC1131513  PMID: 2194451

Abstract

In this paper we present a protocol that allows a dynamic analysis of disulphide-bridge formation, based on freezing the intermediates by acid/acetone precipitation, followed by digestion with pepsin and direct fast-atom-bombardment mass-spectrometric analysis. A rapid definition of the exact nature of disulphide bridges formed can be obtained via a definitive assignment of disulphide-linked peptides according to their unique mass values. With the use of an appropriate thiol concentration, scrambling of the native disulphide bonds in bovine insulin occurs, and the process is catalysed by protein disulphide-isomerase (EC 5.3.4.1). The disruption of native and the formation of new disulphide bonds can be monitored as described above, and interestingly B-chain dimers containing Cys-B7-Cys-B7 and Cys-B7-Cys-B19 bonds are detected.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

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