Skip to main content
Biochemical Journal logoLink to Biochemical Journal
. 1975 Dec;151(3):615–624. doi: 10.1042/bj1510615

Interchain disulphide bridges of mouse immunoglobulin M.

C P Milstein, N E Richardson, E V Dieverson, A Feinstein
PMCID: PMC1172409  PMID: 766753

Abstract

Mouse IgM (immunoglobulin M) was selectively and partially reduced and treated with iodo[2-14C]acetate to label the interchain disulphide bridges. The carboxymethylation was studied in some detail. The labelled peptides were purified, sequenced and positioned by homology with human IgM. Only peptides originating from three interchain disulphide bridges were labelled, in contrast with the four labelled bridges obtained in human IgM under the same conditions. These peptides are homologous to human bridge peptides forming the heavy-light bridge and two inter-heavy bridges, one present in the CMU2 region and the other in the C-terminal region. The inter-heavy bridge in the Cmu2 region was alone cleaved and radioactively labelled in selectively reduced IgM held together as a pentamer by non-covalen interactions. The same bridge was the only one to be totally cleaved in subunits released after more extensive, though still selective, reduction. In the light of these results a possible arrangement of the disulphide bridges of the mouse IgM.

Full text

PDF
615

Images in this article

Selected References

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

  1. Abel C. A., Grey H. M. Carboxy-terminal amino acids of gamma-A and gamma-M heavy chains. Science. 1967 Jun 23;156(3782):1609–1610. doi: 10.1126/science.156.3782.1609. [DOI] [PubMed] [Google Scholar]
  2. Askonas B. A., Parkhouse R. M. Assembly of immunoglobulin M. Blocked thiol groups of intracellular 7S subunits. Biochem J. 1971 Jul;123(4):629–634. doi: 10.1042/bj1230629. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Beale D., Buttress N. Studies on a human 19-S immunoglobulin M. The arrangement of inter-chain disulphide bridges and carbohydrate sites. Biochim Biophys Acta. 1969 May;181(1):250–267. doi: 10.1016/0005-2795(69)90248-7. [DOI] [PubMed] [Google Scholar]
  4. Beale D., Feinstein A. Evidence for C-terminal intra-subunit disulphide bridges between immunoglobulin-M heavy chains. FEBS Lett. 1970 Apr 2;7(2):175–176. doi: 10.1016/0014-5793(70)80149-1. [DOI] [PubMed] [Google Scholar]
  5. Beale D., Feinstein A. Studies on the reduction of a human 19S immunoglobulin M. Biochem J. 1969 Apr;112(2):187–194. doi: 10.1042/bj1120187. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. CHAPLIN H., COHEN S., PRESS E. M. PREPARATION AND PROPERTIES OF THE PEPTIDE CHAINS OF NORMAL HUMAN 19 S GAMMA-GLOBULIN (IGM). Biochem J. 1965 Apr;95:256–261. doi: 10.1042/bj0950256. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Chuang C. Y., Capra J. D., Kehoe J. M. Evolutionary relationship between carboxyterminal region of a human alpha chain and other immunoglobulin heavy chain constant regions. Nature. 1973 Jul 20;244(5412):158–160. doi: 10.1038/244158a0. [DOI] [PubMed] [Google Scholar]
  8. FLEISCHMAN J. B., PORTER R. R., PRESS E. M. THE ARRANGEMENT OF THE PEPTIDE CHAINS IN GAMMA-GLOBULIN. Biochem J. 1963 Aug;88:220–228. doi: 10.1042/bj0880220. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Frangione B., Milstein C. Variations in the S-S bridges of immunoglobins G: interchain disulfide bridges of gamma G3 myeloma proteins. J Mol Biol. 1968 May 14;33(3):893–906. doi: 10.1016/0022-2836(68)90326-4. [DOI] [PubMed] [Google Scholar]
  10. Frangione B., Prelli F., Mihaesco C., Franklin E. C. Interchain and intrachain disulfide bridges of a human immunoglobulin M: detection of a unique fragment. Proc Natl Acad Sci U S A. 1971 Jul;68(7):1547–1551. doi: 10.1073/pnas.68.7.1547. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. GUNDLACH H. G., MOORE S., STEIN W. H. The reaction of iodoacetate with methionine. J Biol Chem. 1959 Jul;234(7):1761–1764. [PubMed] [Google Scholar]
  12. Kownatzki E. Disulfide bonds of human IgM: differential sensitivity to reductive cleavage. Scand J Immunol. 1973;2(4):433–437. doi: 10.1111/j.1365-3083.1973.tb02052.x. [DOI] [PubMed] [Google Scholar]
  13. McIntire K. R., Asofsky R. M., Potter M., Kuff E. L. Macroglobulin-producing plasma-cell tumor in mice: identification of a new light chain. Science. 1965 Oct 15;150(3694):360–363. [PubMed] [Google Scholar]
  14. Mendez E., Prelli F., Frangione B., Franklin E. C. Characterization of a disulfide bridge linking the J chain to the alpha chain of polymeric immunoglobulin A. Biochem Biophys Res Commun. 1973 Dec 19;55(4):1291–1297. doi: 10.1016/s0006-291x(73)80034-8. [DOI] [PubMed] [Google Scholar]
  15. Mestecky J., Schrohenloher R. E., Kulhavy R., Wright G. P., Tomana M. Site of J chain attachment to human polymeric IgA. Proc Natl Acad Sci U S A. 1974 Feb;71(2):544–548. doi: 10.1073/pnas.71.2.544. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Mestecky J., Schrohenloher R. E. Site of attachment of J chain to human immunoglobulin M. Nature. 1974 Jun 14;249(458):650–652. doi: 10.1038/249650a0. [DOI] [PubMed] [Google Scholar]
  17. Mestecky J., Zikan J., Butler W. T., Kulhavy R. Studies on human secretory immunoglobulin A. 3. J chain. Immunochemistry. 1972 Sep;9(9):883–890. doi: 10.1016/0019-2791(72)90162-0. [DOI] [PubMed] [Google Scholar]
  18. Metzger H. Structure and function of gamma M macroglobulins. Adv Immunol. 1970;12:57–116. doi: 10.1016/s0065-2776(08)60168-6. [DOI] [PubMed] [Google Scholar]
  19. Milstein C. P., Feinstein A. Comparative studies of two types of bovine immunoglobulin G heavy chains. Biochem J. 1968 Apr;107(4):559–564. doi: 10.1042/bj1070559. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Milstein C. P., Milstein C. Glycopeptides from human kappa-chains. Biochem J. 1971 Jan;121(2):211–215. doi: 10.1042/bj1210211. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Parkhouse R. M., Askonas B. A., Dourmashkin R. R. Electron microscopic studies of mouse immunoglobulin M; structure and reconstitution following reduction. Immunology. 1970 Apr;18(4):575–584. [PMC free article] [PubMed] [Google Scholar]
  22. Parkhouse R. M. Non-covalent association of IgM subunits produced by reduction and alkylation. Immunology. 1974 Dec;27(6):1063–1071. [PMC free article] [PubMed] [Google Scholar]
  23. Poljak R. J., Amzel L. M., Avey H. P., Chen B. L., Phizackerley R. P., Saul F. Three-dimensional structure of the Fab' fragment of a human immunoglobulin at 2,8-A resolution. Proc Natl Acad Sci U S A. 1973 Dec;70(12):3305–3310. doi: 10.1073/pnas.70.12.3305. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Putnam F. W., Florent G., Paul C., Shinoda T., Shimizu A. Complete amino acid sequence of the Mu heavy chain of a human IgM immunoglobulin. Science. 1973 Oct 19;182(4109):287–291. doi: 10.1126/science.182.4109.287. [DOI] [PubMed] [Google Scholar]
  25. ROSEVEAR J. W., SMITH E. L. Glycopeptides. I. Isolation and properties of glycopeptides from a fraction of human gamma-globulin. J Biol Chem. 1961 Feb;236:425–435. [PubMed] [Google Scholar]
  26. Robinson E. A., Appella E., McIntire K. R. Structural studies of a heavy chain from a mouse immunoglobulin M with antibody activity. J Biol Chem. 1973 Oct 25;248(20):7112–7117. [PubMed] [Google Scholar]
  27. Shimizu A., Putnam F. W., Paul C., Clamp J. R., Johnson I. Structure and role of the five glycopeptides of human IgM immunoglobulins. Nat New Biol. 1971 May 19;231(20):73–76. doi: 10.1038/newbio231073a0. [DOI] [PubMed] [Google Scholar]
  28. 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]
  29. Tomasi T. B., Jr Production of a noncovalently bonded pentamer of immunoglobulin M: relationship of J chain. Proc Natl Acad Sci U S A. 1973 Dec;70(12):3410–3414. doi: 10.1073/pnas.70.12.3410. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Watanabe S., Barnikol H. U., Horn J., Bertram J., Hilschmann N. Die Primärstruktur eines monoklonalen IgM-Immunoglobulins (Makroglobulin Gal.), II. Die Aminosäuresequenz der H-Kette (mu-Typ,Subgruppe HIII), Struktur des gesamten IgM-Moleküls. Hoppe Seylers Z Physiol Chem. 1973 Oct-Nov;354(10-11):1505–1509. [PubMed] [Google Scholar]
  31. Woods K. R., Wang K. T. Separation of dansyl-amino acids by polyamide layer chromatography. Biochim Biophys Acta. 1967 Feb 21;133(2):369–370. doi: 10.1016/0005-2795(67)90078-5. [DOI] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES