Skip to main content
NCBI home page
Immunology logoLink to Immunology
. 1988 Oct;65(2):259–265.

The rat T-cell differentiation marker RT6.1 is more polymorphic than its alloantigenic counterpart RT6.2.

F Koch 1, A Kashan 1, H G Thiele 1
PMCID: PMC1384922  PMID: 3263942

Abstract

Utilizing allotype-specific antibodies to immunoprecipitate RT6.2 from DA.6B rat lymphocyte lysates, we have shown this antigen recently to be composed of two related, non-glycosylated polypeptides with apparent molecular weights (MW) of 24,000 and 26,000 (reducing conditions), which evidently are anchored in the cell membrane by covalent linkage to phosphatidylinositol. The present report shows that RT6.1 allotype-specific antibodies precipitate a more complex pattern of bands from LEW.6A rat lymphocyte lysates. These consist of an endo-F-resistant RT6.2-like 25/27,000 MW doublet (reducing conditions) as well as at least five additional endo-F-sensitive, endo-H-resistant polypeptides of 30,000, 32,000, 33,000, 34,000, 35,000 MW. Endo-F treatment seems to convert the additional higher to the lower molecular weight forms. In contrast to the endo-F-resistant 25/27,000 MW doublet, the higher MW forms of RT6.1 partly bind to lentil lectin and concanavalin A (Con A). Two-dimensional electrophoretic analyses (NEPHGE/SDS-PAGE) reveal similar patterns of charge heterogeneity of the lower and higher MW forms of RT6.1. Neuraminidase treatment does not affect the pIs of the lower MW forms but shifts the pIs of the higher MW forms to those of the lower ones. All forms of RT6.1 evidently employ the covalent linkage to phosphatidylinositol for membrane anchorage. Identical patterns of molecular forms--the doublet for RT6.2, the polymorphic pattern for RT6.1--are observed upon immunoprecipitation of the alloantigens from the lysates of corresponding series of inbred strains of rats with both allotype-specific antibodies and a polyclonal rabbit serum recognizing a common determinant on both alloantigens.

Full text

PDF
259

Images in this article

261Figure 1
on p.261
262Figure 2
on p.262
262Figure 3
on p.262
262Figure 4
on p.262
263Figure 5
on p.263
263Figure 6
on p.263

Selected References

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

  1. Borst J., Alexander S., Elder J., Terhorst C. The T3 complex on human T lymphocytes involves four structurally distinct glycoproteins. J Biol Chem. 1983 Apr 25;258(8):5135–5141. [PubMed] [Google Scholar]
  2. Breitbart R. E., Andreadis A., Nadal-Ginard B. Alternative splicing: a ubiquitous mechanism for the generation of multiple protein isoforms from single genes. Annu Rev Biochem. 1987;56:467–495. doi: 10.1146/annurev.bi.56.070187.002343. [DOI] [PubMed] [Google Scholar]
  3. Carlsson S. R., Stigbrand T. Partial characterization of the oligosaccharides of mouse thymocyte Thy-1 glycoprotein. Biochem J. 1984 Jul 15;221(2):379–392. doi: 10.1042/bj2210379. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Cleveland D. W., Fischer S. G., Kirschner M. W., Laemmli U. K. Peptide mapping by limited proteolysis in sodium dodecyl sulfate and analysis by gel electrophoresis. J Biol Chem. 1977 Feb 10;252(3):1102–1106. [PubMed] [Google Scholar]
  5. DeWitt C. W., MCCullough M. Ag-F: serological and genetic identification of a new locus in the rat governing lymphocyte membrane antigens. Transplantation. 1975 Apr;19(4):310–317. [PubMed] [Google Scholar]
  6. Gahmberg C. G. Molecular characterization of the human red cell Rho(D) antigen. EMBO J. 1983;2(2):223–227. doi: 10.1002/j.1460-2075.1983.tb01409.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Greiner D. L., Handler E. S., Nakano K., Mordes J. P., Rossini A. A. Absence of the RT-6 T cell subset in diabetes-prone BB/W rats. J Immunol. 1986 Jan;136(1):148–151. [PubMed] [Google Scholar]
  8. Greiner D. L., Mordes J. P., Handler E. S., Angelillo M., Nakamura N., Rossini A. A. Depletion of RT6.1+ T lymphocytes induces diabetes in resistant biobreeding/Worcester (BB/W) rats. J Exp Med. 1987 Aug 1;166(2):461–475. doi: 10.1084/jem.166.2.461. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Koch F., Thiele H. G., Low M. G. Release of the rat T cell alloantigen RT-6.2 from cell membranes by phosphatidylinositol-specific phospholipase C. J Exp Med. 1986 Oct 1;164(4):1338–1343. doi: 10.1084/jem.164.4.1338. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Kornfeld R., Kornfeld S. Assembly of asparagine-linked oligosaccharides. Annu Rev Biochem. 1985;54:631–664. doi: 10.1146/annurev.bi.54.070185.003215. [DOI] [PubMed] [Google Scholar]
  11. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  12. Lubaroff D. M., Greiner D. L., Reynolds C. W. Investigations of T-lymphocyte subpopulations in the rat using alloantigenic markers. Transplant Proc. 1979 Mar;11(1):1092–1094. [PubMed] [Google Scholar]
  13. Mage M. G. Separation of lymphocytes on antibody-coated plates. Methods Enzymol. 1984;108:118–124. [PubMed] [Google Scholar]
  14. Marchalonis J. J., Cone R. E., Santer V. Enzymic iodination. A probe for accessible surface proteins of normal and neoplastic lymphocytes. Biochem J. 1971 Oct;124(5):921–927. doi: 10.1042/bj1240921. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Morrison M. H., Chaney W. G., Esselman W. J. Molecular weight and charge heterogeneity of Thy-1 glycoprotein in different populations of T-cells. Mol Immunol. 1984 May;21(5):405–413. doi: 10.1016/0161-5890(84)90038-5. [DOI] [PubMed] [Google Scholar]
  16. 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]
  17. Parekh R. B., Tse A. G., Dwek R. A., Williams A. F., Rademacher T. W. Tissue-specific N-glycosylation, site-specific oligosaccharide patterns and lentil lectin recognition of rat Thy-1. EMBO J. 1987 May;6(5):1233–1244. doi: 10.1002/j.1460-2075.1987.tb02359.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Sheares B. T., Robbins P. W. Glycosylation of ovalbumin in a heterologous cell: analysis of oligosaccharide chains of the cloned glycoprotein in mouse L cells. Proc Natl Acad Sci U S A. 1986 Apr;83(7):1993–1997. doi: 10.1073/pnas.83.7.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Swiedler S. J., Freed J. H., Tarentino A. L., Plummer T. H., Jr, Hart G. W. Oligosaccharide microheterogeneity of the murine major histocompatibility antigens. Reproducible site-specific patterns of sialylation and branching in asparagine-linked oligosaccharides. J Biol Chem. 1985 Apr 10;260(7):4046–4054. [PubMed] [Google Scholar]
  20. Thiele H. G., Arndt R., Stark R., Wonigeit K. Detection and partial molecular characterization of the rat T-lymphocyte surface protein L21 by allo-(anti-RT-Ly-2.2) and xeno-(anti-RT-LN-LyIg) sera. Transplant Proc. 1979 Sep;11(3):1636–1638. [PubMed] [Google Scholar]
  21. Thiele H. G., Koch F., Hamann A., Arndt R. Biochemical characterization of the T-cell alloantigen RT-6.2. Immunology. 1986 Oct;59(2):195–201. [PMC free article] [PubMed] [Google Scholar]
  22. Wonigeit K. Characterization of the RT-Ly-1 and RT-Ly-2 alloantigenic systems by congenic rat strains. Transplant Proc. 1979 Sep;11(3):1631–1635. [PubMed] [Google Scholar]
  23. Wonigeit K., Schwinzer R. Polyclonal activation of rat T lymphocytes by RT6 alloantisera. Transplant Proc. 1987 Feb;19(1 Pt 1):296–299. [PubMed] [Google Scholar]
  24. van der Feltz M. J., Ploegh H. L. Immunochemical analysis of glycosylated and nonglycosylated DLA class I antigens. Immunogenetics. 1984;19(2):95–107. doi: 10.1007/BF00387852. [DOI] [PubMed] [Google Scholar]

Articles from Immunology are provided here courtesy of British Society for Immunology

RESOURCES