Skip to main content
SpringerLink
Log in

Carbohydrate moieties of rat MHC class I antigens

  • Published:
Immunogenetics Aims and scope Submit manuscript

Abstract

The rat major histocompatibility complex class I antigens RT1.Au and RT1.Eu from the u haplotype and RT1.An from the n haplotype were labeled with 14C-asparagine or with 3H-fucose, mannose, galactose, and N-acetylglucosamine. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis showed complete removal of radioactivity from the sugar-labeled antigen heavy chains by digestion with glycopeptidase F, an enzyme that removes N-linked glycans completely. High performance liquid chromatography analysis of the tryptic digests of the mixed sugar-labeled and asparagine-labeled antigens demonstrated that all the sugar-labeled peptides were coincident with asparagine-labeled peptides. The An antigen showed three glycopeptides, each of which had different amounts of sugar radioactivity. The antigens Au and Eu showed two glycopeptides with different amounts of radioactivity but at identical positions in the two antigens. Antigen Eu had an additional glycopeptide with a lower amount of radioactivity. The positions of the glycopeptides from the Au and Eu antigens were different from those of the An antigen. The peptide profiles of the 14C-asparagine-labeled Au and Eu antigens demonstrated distinct differences between the molecules. The results of this study show that: (a) all the glycans on rat class I antigens are N-linked, as they are on H-2 and HLA class I antigens; (b) there are compositional differences among the glycans in each of the three antigens; (c) the glycosylation pattern of the rat class I antigens is similar to that of the mouse class I antigens, which contain two or three glycans, in contrast to that of the human class I antigens, which contain only one glycan; and (d) the antigens Au and Eu from the same haplotype are more closely related to each other than they are to the An antigen.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Blankenhorn, E. P. and Cramer, D. V.: Orientation of the loci encoding RT1.B polypeptides in the major histocompatibility complex of the rat. Immunogenetics 21: 135–142, 1985

    Google Scholar 

  • Cortese Hassett, A. L., Stranick, K. S., Locker, J., Kunz, H. W., and Gill, T. J., III: Molecular analysis of the rat MHC. I. Delineation of the major regions in the MHC and the grc. J. Immunol. 137: 373–378, 1986

    Google Scholar 

  • Cowing, C. and Chapdelaine, J. M.: T cells discriminate between Ia antigens expressed on allogeneic accessory cells and B cells: A potential function for carbohydrate side chains on la molecules. Proc. Natl. Acad. Sci. U.S.A. 80: 6000–6004, 1983

    Google Scholar 

  • Cullen, S. E., Kindle, C. S., Shreffler, D. C., and Cowing, C.: Differential glycosylation of murine B cell and spleen adherent cell Ia antigens. J. Immunol. 127: 1478–1484, 1981

    Google Scholar 

  • Freed, J. H. and Nathenson, S. G.: Similarity of the carbohydrate structures of H-2 and la glycoproteins. J. Immunol. 119: 477–482, 1977

    Google Scholar 

  • Gill, T. J., III, Cramer, D. V., Kunz, H. W., and Misra, D. N.: Structure and function of the major histocompatibility complex of the rat. J. Immunogenet. 10: 261–273, 1983

    Google Scholar 

  • Gill, T. J., III, Kunz, H. W., Misra, D. N., and Cortese Hassett, A. L.: The major histocompatibility complex of the rat. Transplantation, in press, 1987

  • Günther, E. and Wurst, W.: Cytotoxic T lymphocytes of the rat are predominantly restricted by RT1.A and not RT1.C-determined major histocompatibility class I antigens. Immunogenetics 20: 1–12, 1984

    Google Scholar 

  • Haustein, D. and Günther, E.: Biochemical analysis of gene products of major histocompatibility recombinant haplotypes in the rat. Eur. J. Immunol. 10: 615–620, 1980

    Google Scholar 

  • Haustein, D., Stock, W., and Günther, E.: Rat major histocompatibility RT1. C antigens of restricted tissue distribution consist of two polypeptide chains with molecular weights of about 42 000 and 12 500. Immunogenetics 15: 271–277, 1982

    Google Scholar 

  • Howard, J. C.: The major histocompatibility complex of the rat: A partial review. Metabolism 32: 41–50, 1983

    Google Scholar 

  • Hubbard, S. C. and Ivatt, R. J.: Synthesis and processing of asparaginelinked oligosaccharides. Annu. Rev. Biochem. 50: 555–583, 1981

    Google Scholar 

  • Inomata, T., Kunz, H. W., and Gill, T. J., III: The rat Tla-region alloantigens. Transplant. Proc., in press, 1987

  • Kimball, E. S. and Coligan, J. E.: Structure of class I major histocompatibility antigens. Contemp. Top. Mol. Immunol. 9: 1–63, 1983

    Google Scholar 

  • Kimball, E. S., Maloy, W. L., and Coligan, J. E.: Evidence for three carbohydrate prosthetic groups on mouse major histocompatibility antigens H-2Kd and H-2Db. Mol. Immunol. 18: 677–680, 1981

    Google Scholar 

  • Kunz, H. W., Gill, T. J., III, and Misra, D. N.: The identification and mapping of a second class I locus in the major histocompatibility complex of the rat. J. Immunol. 128: 402–408, 1982

    Google Scholar 

  • Kvist, S., Sandberg-Tragardh, L., Ostberg, L., and Peterson, P. A.: Isolation and partial characterization of papain-solubilized murine H-2 antigens. Biochemistry 16: 4415–4420, 1977

    Google Scholar 

  • Lafay, F., Ewald, S. J., McMillan, M., Frelinger, J. A., and Hood, L.: Tryptic peptide map analyses of mouse transplantation antigens. Immunogenetics 12: 21–32, 1981

    Google Scholar 

  • Landolfi, N. F. and Cook, R. G.: Activated T-lymphocytes express class I molecules which are hyposialylated compared to other lymphocyte populations. Mol. Immunol. 23: 297–309, 1986

    Google Scholar 

  • Landolfi, N. F., Rich, R. R., and Cook, R. G.: The Qa-1 alloantigens. III. Biochemical analysis of the structure and extent of polymorphism of the Qa-1 allelic products. J. Immunol. 135: 1264–1270, 1985

    Google Scholar 

  • Le, A. V. and Doyle, D.: Differential regulation of mouse H-2 alloantigens. Biochemistry 21: 5738–5744, 1982

    Google Scholar 

  • Lillehoj, E. P., Myers, N. B., Lee, D. R., Hansen, T. H., and Coligan, J. E.: Structural definition of a family of Ld-like molecules distributed among four of seven haplotypes compared. J. Immunol. 135: 1271–1275, 1985

    Google Scholar 

  • Misra, D. N., Kunz, H. W., and Gill, T. J., III: Analysis of class I MHC antigens in the rat by monoclonal antibodies. J. Immunol. 134: 2520–2528, 1985

    Google Scholar 

  • Misra, D. N., Kunz, H. W., Cortese Hassett, A. L., and Gill, T. J., III: Comparison of rat MHC class I antigens by peptide mapping. Immunogenetics 25: 35–46, 1987a

    Google Scholar 

  • Misra, D. N., Kunz, H. W., Cortese Hassett, A. L., and Gill, T. J., III: Comparison of rat MHC class I antigens by peptide mapping. Transplant. Proc., in press, 1987b

  • Miyazaki, J.-I., Appella, E., Zhao, H., Forman, J., and Ozato, K.: Expression and function of a nonglycosylated major histocompatibility class I antigen. J. Exp. Med. 163: 856–871, 1986

    Google Scholar 

  • Mölders, H. H., Breuning, M. H., Ivanyi, P., and Ploegh, H. L.: Biochemical analysis of variant HLA-B27 antigen. Hum. Immunol. 6: 111–117, 1983

    Google Scholar 

  • Nathenson, S. G. and Muramatsu, T.: Properties of the carbohydrate portion of mouse H-2 alloantigen glycoproteins. In G. A. Jemieson and T. J. Greenwalt (eds.): Glycoproteins of Blood Cells and Plasma, pp. 245–262, Lippincott, Philadelphia, 1971

    Google Scholar 

  • Nilsson, S. F. and Wigzell, H.: Isolation and partial characterization of rat lymphoid cell surface histocompatibility antigens and immunoglobulins. Scand. J. Immunol. 7: 307–314, 1978

    Google Scholar 

  • Parham, P., Humphreys, R. E., Turner, M. J., and Strominger, J. L.: Heterogeneity of HL-A preparations is due to variable sialic acid content. Proc. Natl. Acad. Sci. U.S.A. 71: 3998–4001, 1974

    Google Scholar 

  • Parham, P., Alpert, B. N., Orr, H. T., and Strominger, J. L.: Carbohydrate moiety of HLA antigens: Antigenic properties and amino acid sequences around the site of glycosylation. J. Biol. Chem. 252: 7555–7567, 1977

    Google Scholar 

  • Pearson, T. W. and Anderson, N. L.: Use of high-resolution two-dimensional gel electrophoresis for analysis of monoclonal antibodies and their specific antigens. Methods Enzymol. 92: 196–220, 1983

    Google Scholar 

  • Porzio, M. A. and Pearson, A. M.: Improved resolution of myofibrillar proteins with sodium dodecyl-polyacrylamide gel electrophoresis. Biochim. Biophys. Acta 490: 27–34, 1977

    Google Scholar 

  • Rothenberg, E. and Boyse, E. A.: Synthesis and processing of molecules bearing thymus leukemia antigen. J. Exp. Med. 150: 777–791, 1979

    Google Scholar 

  • Rothenberg, E. and Triglia, D.: Structure and expression of glycoproteins controlled by the Qa-1 aallele. Immunogenetics 14: 455–468, 1981

    Google Scholar 

  • Sporer, R., Mason, L. A., and Götze, D.: A biochemical analysis of the membrane-associated gene products of major histocompatibility complex of the rat. J. Immunol. 122: 2162–2166, 1979

    Google Scholar 

  • Stock, W. and Günther, E.: Serologic and cellular characterization of products of a new major histocompatibility gene region, RT1.C, of the rat; possible homology to mouse H-2 Qa. J. Immunol. 128: 1923–1928, 1982

    Google Scholar 

  • Strominger, J. L., Engelhard, V. H., Fuks, A., Guild, B. C., Hyafil, F., Kaufman, J. F., Korman, A. J., Kostyk, T. G., Krangel, M. S., Lancet, D., Lopez de Castro, J. A., Mann, D. L., Orr, H. T., Parham, P. R., Parker, K. C., Ploegh, H. L., Pober, J. S., Robb, R. J., and Shackelford, D. A.: Biochemical analysis of products of the MHC. In M. E. Dorf (ed.): The Role of the Major Histocompatibility Complex in Immunobiology, pp. 115–172, Garland Publishing, Inc., New York, 1981

    Google Scholar 

  • Tarentino, A. L., Gomez, C. M., and Plummer, T. H., Jr.: Deglycosylation of asparagine-linked by peptide: N-glycosidase F. Biochemistry 24: 4665–4671, 1985

    Google Scholar 

  • Terhorst, C., Parham, P., Mann, D. L., and Strominger, J. L.: Structure of HLA antigens: Amino acids and carbohydrate compositions and NH2- terminal sequences of four antigenic preparations. Proc. Natl. Acad. Sci. U.S.A. 73: 910–914, 1976

    Google Scholar 

  • van der Feltz, M. J. M. and Ploegh, H. L.: Immunochemical analysis of glycosylated and nonglycosylated DLA class I antigens. Immunogenetics 19: 95–107, 1984

    Google Scholar 

  • Van de Rijn, M., Lerch, P. G., Knowles, R. W., and Terhorst, C.: The thymic differentiation markers T6 and M241 are two unusual MHC class I antigens. J. Immunol. 131: 851–855, 1983

    Google Scholar 

  • Widacki, S. M., Flaherty, L., and Cook, R. G.: Biochemical characterization of the molecules reactive with Qa-6 antiserum and monoclonal antibody 20-8-4: Evidence for structural similarity with the Qa-2 molecule. J. Immunol. 135: 3333–3339, 1985

    Google Scholar 

  • Willoughby, E. M., Turner, M. J., and Sanderson, A. R.: Incorporation of rat histocompatibility (AgB) antigens into liposomes, and their susceptibility to immune lysis. Eur. J. Immunol. 8: 628–634, 1978

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Pathology, University of Pittsburgh School of Medicine, 15261, Pittsburgh, PA, USA

    Dhirendra N. Misra, Heinz W. Kunz & Thomas J. Gill III

Authors
  1. Dhirendra N. Misra
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Heinz W. Kunz
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Thomas J. Gill III
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Misra, D.N., Kunz, H.W. & Gill, T.J. Carbohydrate moieties of rat MHC class I antigens. Immunogenetics 26, 204–210 (1987). https://doi.org/10.1007/BF00346513

Download citation

  • Received:

  • Revised:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00346513

Keywords

Search

Navigation