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. 1975 Aug;72(8):3029–3033. doi: 10.1073/pnas.72.8.3029

A rapid and specific method for isolation of thiol-containing peptides from large proteins by thiol-disulfide exchange on a solid support.

T A Egorov, A Svenson, L Rydén, J Carlsson
PMCID: PMC432912  PMID: 1059090

Abstract

Activated thiol-Sepharose [agarose-(glutathione-2-pyridyl disulfide) conjugate] has been used to immobilize proteins with a single or a few thiol groups via disulfide bridges. The immobilized proteins were subsequently proteolytically degraded. After washing, the thiol-containing peptides were eluted with a reducing agent. A single preparative paper electrophoresis, occasionally after a modification such as oxidation, was sufficient to obtain pure peptides in good yields. The method was applied to the major parvalbumin from hake muscle (a protein with 108 amino acid residues and one cysteine residue), to mercaptalbumin from bovine serum (565 residues and one cysteine), and to human serum ferroxidase [EC 1.16.3.1; iron (II):oxygen oxidoreductase] (ceruloplasmin) (1065 residues and three cysteines). The use of the technique, e.g., as a simple means of obtaining homologous peptides in related proteins, is discussed.

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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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