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. 1983 Dec 1;158(6):1979–1992. doi: 10.1084/jem.158.6.1979

Identification of a new component in the murine Ia molecular complex

PMCID: PMC2187163  PMID: 6580373

Abstract

In this report, we describe a previously unidentified component in the murine Ia antigen complex. SDS-PAGE analysis of anti-Ia immunoprecipitates prepared from spleen cells biosynthetically labeled with 35S-sulfate showed no detectable incorporation of 35SO4 into alpha, beta, or Ii chains but did not reveal the presence of a novel sulfate-bearing molecule of considerable molecular weight heterogeneity (46-69-kdaltons). The 46-69-kdalton molecule could be precipitated with monoclonal antibodies specific for I-A, I-E, and Ii glycoproteins but was not seen in control precipitates, nor in association with IgG or class I MHC molecules. Preliminary biochemical characterization indicated that the 46-69-kdalton product is extremely polydisperse, both in charge and apparent molecular weight, is sensitive to proteases, and bears the sulfate moiety on a large pronase-resistant structure. These results suggested this component might be a proteoglycan. Definitive identification of this component as a proteoglycan was accomplished by selective enzymatic degradation experiments which showed that the sulfate-bearing component of the 46- 69-kdalton molecule is chondroitin 6-sulfate.

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Selected References

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  1. Auffray C., Korman A. J., Roux-Dosseto M., Bono R., Strominger J. L. cDNA clone for the heavy chain of the human B cell alloantigen DC1: strong sequence homology to the HLA-DR heavy chain. Proc Natl Acad Sci U S A. 1982 Oct;79(20):6337–6341. doi: 10.1073/pnas.79.20.6337. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Baenziger J., Kornfeld S. Structure of the carbohydrate units of IgA1 immunoglobulin. II. Structure of the O-glycosidically linked oligosaccharide units. J Biol Chem. 1974 Nov 25;249(22):7270–7281. [PubMed] [Google Scholar]
  3. Benacerraf B., Germain R. N. The immune response genes of the major histocompatibility complex. Immunol Rev. 1978;38:70–119. doi: 10.1111/j.1600-065x.1978.tb00385.x. [DOI] [PubMed] [Google Scholar]
  4. Benacerraf B., McDevitt H. O. Histocompatibility-linked immune response genes. Science. 1972 Jan 21;175(4019):273–279. doi: 10.1126/science.175.4019.273. [DOI] [PubMed] [Google Scholar]
  5. Benoist C. O., Mathis D. J., Kanter M. R., Williams V. E., 2nd, McDevitt H. O. The murine Ia alpha chains, E alpha and A alpha, show a surprising degree of sequence homology. Proc Natl Acad Sci U S A. 1983 Jan;80(2):534–538. doi: 10.1073/pnas.80.2.534. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Bhavanandan V. P., Umemoto J., Banks J. R., Davidson E. A. Isolation and partial characterization of sialoglycopeptides produced by a murine melanoma. Biochemistry. 1977 Oct 4;16(20):4426–4437. doi: 10.1021/bi00639a016. [DOI] [PubMed] [Google Scholar]
  7. Cullen S. E., Kindle C. S., Shreffler D. C., Cowing C. Differential glycosylation of murine B cell and spleen adherent cell Ia antigens. J Immunol. 1981 Oct;127(4):1478–1484. [PubMed] [Google Scholar]
  8. Cullen S. E., Schwartz B. D. An improved method for isolation of H-2 and Ia alloantigens with immunoprecipitation induced by protein A-bearing staphylococci. J Immunol. 1976 Jul;117(1):136–142. [PubMed] [Google Scholar]
  9. Day C. E., Jones P. P. The gene encoding the Ia antigen-associated invariant chain (Ii) is not linked to the H-2 complex. Nature. 1983 Mar 10;302(5904):157–159. doi: 10.1038/302157a0. [DOI] [PubMed] [Google Scholar]
  10. Drapeau G. R., Boily Y., Houmard J. Purification and properties of an extracellular protease of Staphylococcus aureus. J Biol Chem. 1972 Oct 25;247(20):6720–6726. [PubMed] [Google Scholar]
  11. Freed J. H., Nathenson S. G. Similarity of the carbohydrate structures of H-2 and Ia glycoproteins. J Immunol. 1977 Aug;119(2):477–482. [PubMed] [Google Scholar]
  12. Hauptfeld V., Braciale T. J., Shreffler D. C. Differences in expression of MHC products between several H-2-restricted CTL clones. J Immunol. 1982 May;128(5):2026–2031. [PubMed] [Google Scholar]
  13. Heifetz A., Kinsey W. H., Lennarz W. J. Synthesis of a novel class of sulfated glycoproteins in embryonic liver and lung. J Biol Chem. 1980 May 25;255(10):4528–4534. [PubMed] [Google Scholar]
  14. Heifetz A., Watson C., Johnson A. R., Roberts M. K. Sulfated glycoproteins secreted by human vascular endothelial cells. J Biol Chem. 1982 Nov 25;257(22):13581–13586. [PubMed] [Google Scholar]
  15. Hood L., Steinmetz M., Malissen B. Genes of the major histocompatibility complex of the mouse. Annu Rev Immunol. 1983;1:529–568. doi: 10.1146/annurev.iy.01.040183.002525. [DOI] [PubMed] [Google Scholar]
  16. Hortin G., Natowicz M., Pierce J., Baenziger J., Parsons T., Boime I. Metabolic labeling of lutropin with [35S]sulfate. Proc Natl Acad Sci U S A. 1981 Dec;78(12):7468–7472. doi: 10.1073/pnas.78.12.7468. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Huttner W. B. Sulphation of tyrosine residues-a widespread modification of proteins. Nature. 1982 Sep 16;299(5880):273–276. doi: 10.1038/299273a0. [DOI] [PubMed] [Google Scholar]
  18. Jones P. P. Analysis of H-2 and Ia molecules by two-dimensional gel electrophoresis. J Exp Med. 1977 Nov 1;146(5):1261–1279. doi: 10.1084/jem.146.5.1261. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Kappler J. W., Skidmore B., White J., Marrack P. Antigen-inducible, H-2-restricted, interleukin-2-producing T cell hybridomas. Lack of independent antigen and H-2 recognition. J Exp Med. 1981 May 1;153(5):1198–1214. doi: 10.1084/jem.153.5.1198. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Karr R. W., Kannapell C. C., Stein J. A., Gebel H. M., Mann D. L., Duquesnoy R. J., Fuller T. C., Rodey G. E., Schwartz B. D. Molecular relationships of the human B cell alloantigens, MT2, MB3, MT4, and DR5. J Immunol. 1982 Apr;128(4):1809–1818. [PubMed] [Google Scholar]
  21. Kimata K., Okayama M., Ooira A., Suzuki S. Heterogeneity of proteochondroitin sulfates produced by chondrocytes at different stages of cytodifferentiation. J Biol Chem. 1974 Mar 10;249(5):1646–1653. [PubMed] [Google Scholar]
  22. Koch N., Hämmerling G. J., Szymura J., Wabl M. R. Ia associated Ii chain is not encoded by chromosome 17 of the mouse. Immunogenetics. 1982;16(6):603–606. doi: 10.1007/BF00372029. [DOI] [PubMed] [Google Scholar]
  23. Koch N., Koch S., Hämmerling G. J. Ia invariant chain detected on lymphocyte surfaces by monoclonal antibody. Nature. 1982 Oct 14;299(5884):644–645. doi: 10.1038/299644a0. [DOI] [PubMed] [Google Scholar]
  24. Lerner E. A., Matis L. A., Janeway C. A., Jr, Jones P. P., Schwartz R. H., Murphy D. B. Monoclonal antibody against an Ir gene product? J Exp Med. 1980 Oct 1;152(4):1085–1101. doi: 10.1084/jem.152.4.1085. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Lin C. C., Rosenthal A. S., Passmore H. C., Hansen T. H. Selective loss of antigen-specific Ir gene function in IA mutant B6.C-H-2bm12 is an antigen presenting cell defect. Proc Natl Acad Sci U S A. 1981 Oct;78(10):6406–6410. doi: 10.1073/pnas.78.10.6406. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. O'Farrell P. H. High resolution two-dimensional electrophoresis of proteins. J Biol Chem. 1975 May 25;250(10):4007–4021. [PMC free article] [PubMed] [Google Scholar]
  27. Rosenthal A. S., Shevach E. M. Function of macrophages in antigen recognition by guinea pig T lymphocytes. I. Requirement for histocompatible macrophages and lymphocytes. J Exp Med. 1973 Nov 1;138(5):1194–1212. doi: 10.1084/jem.138.5.1194. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Yamagata T., Saito H., Habuchi O., Suzuki S. Purification and properties of bacterial chondroitinases and chondrosulfatases. J Biol Chem. 1968 Apr 10;243(7):1523–1535. [PubMed] [Google Scholar]
  29. Yonekura H., Oguri K., Nakazawa K., Shimizu S., Nakanishi Y., Okayama M. Isolation and partial characterization of sulfated glycoproteins synthesized by corneal epithelium. J Biol Chem. 1982 Sep 25;257(18):11166–11175. [PubMed] [Google Scholar]

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