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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.

  1. ASKONAS B. A. A study on globulin formation by plasma-cell neoplasm (5563) transplantable in mice. Biochem J. 1961 Apr;79:33–43. doi: 10.1042/bj0790033. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Awdeh Z. L., Askonas B. A., Williamson A. R. The homogeneous-gamma-G-immunoglobulin produced by mouse plasmacytoma 5563 and its subsequent heterogeneity in serum. Biochem J. 1967 Feb;102(2):548–553. doi: 10.1042/bj1020548. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. HERMANS P. E., McGUCKIN W. F., McKENZIE B. F., BAYRD E. D. Electrophoretic studies of serum proteins in cyanogum gel. Proc Staff Meet Mayo Clin. 1960 Dec 21;35:792–807. [PubMed] [Google Scholar]
  4. Maizel J. V., Jr Acrylamide-gel electrophorograms by mechanical fractionation: radioactive adenovirus proteins. Science. 1966 Feb 25;151(3713):988–990. doi: 10.1126/science.151.3713.988. [DOI] [PubMed] [Google Scholar]
  5. POTTER M., FAHEY J. L., PILGRIM H. I. Abnormal serum protein and bone destruction in transmissible mouse plasma cell neoplasm (multiple myeloma). Proc Soc Exp Biol Med. 1957 Feb;94(2):327–333. doi: 10.3181/00379727-94-22035. [DOI] [PubMed] [Google Scholar]
  6. POTTER M., KUFF E. L. DISORDERS IN THE DIFFERENTIATON OF PROTEIN SECRETION IN NEOPLASTIC PLASMA CELLS. J Mol Biol. 1964 Aug;9:537–544. doi: 10.1016/s0022-2836(64)80225-4. [DOI] [PubMed] [Google Scholar]
  7. Shaeffer J. R. Evidence for soluble alpha-chains as intermediates in hemoglobin synthesis in the rabbit reticulocyte. Biochem Biophys Res Commun. 1967 Aug 23;28(4):647–652. doi: 10.1016/0006-291x(67)90363-4. [DOI] [PubMed] [Google Scholar]
  8. Shapiro A. L., Scharff M. D., Maizel J. V., Jr, Uhr J. W. Polyribosomal synthesis and assembly of the H and L chains of gamma globulin. Proc Natl Acad Sci U S A. 1966 Jul;56(1):216–221. doi: 10.1073/pnas.56.1.216. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Summers D. F., Maizel J. V., Jr, Darnell J. E., Jr Evidence for virus-specific noncapsid proteins in poliovirus-infected HeLa cells. Proc Natl Acad Sci U S A. 1965 Aug;54(2):505–513. doi: 10.1073/pnas.54.2.505. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Williamson A. R., Adkonas B. A. Time course of labeling of light and heavy chains of immunoglobulin G and the turnover of the free light-chain intermediates. Arch Biochem Biophys. 1968 May;125(2):401–409. doi: 10.1016/0003-9861(68)90596-1. [DOI] [PubMed] [Google Scholar]
  11. Williamson A. R., Askonas B. A. Biosynthesis of immunoglobulins: the separate classes of polyribosomes synthesizing heavy and light chains. J Mol Biol. 1967 Jan 28;23(2):201–216. doi: 10.1016/s0022-2836(67)80027-5. [DOI] [PubMed] [Google Scholar]
  12. Williamson A. R., Askonas B. A. Differential reduction of interchain disulphide bonds of mouse immunoglobulin G. Biochem J. 1968 May;107(6):823–828. doi: 10.1042/bj1070823. [DOI] [PMC free article] [PubMed] [Google Scholar]

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