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. 1968 Oct;109(4):637–643. doi: 10.1042/bj1090637

Interchain disulphide-bond formation in the assembly of immunoglobulin G. Heavy-chain dimer as an intermediate

Brigitte A Askonas 1, Alan R Williamson 1
PMCID: PMC1186949  PMID: 5683511

Abstract

1. The role of disulphide-bond formation in the assembly of G2a myeloma protein 5563 was studied by pulse-labelling ascitic plasma cells of tumour-line 5563 for 2–8min. with radioactive amino acids, and analysing the intracellular proteins. Myeloma-protein determinants were first purified by ion-exchange chromatography under conditions that do not dissociate non-covalently linked sub-units of immunoglobulin G. The pulse-labelled material was then analysed by electrophoresis on polyacrylamide gels in sodium dodecyl sulphate–phosphate–urea buffer, which dissociates non-covalently linked sub-units; after gel electrophoresis, radioactive protein bands were located by radioautography, and characterized immunologically after elution. 2. Two heavy-chain intermediates were detected: (i) heavy-chain dimer; (ii) the dimer with one light chain attached. Free light chains had previously been shown to be intermediates in assembly. No evidence for the presence of half-molecules (one light chain attached to one heavy chain) was obtained. The formation of the disulphide bond between the heavy chains thus appears to precede the light-chain–heavy-chain linkage in immunoglobulin G assembly.

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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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