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. 1990 Sep;87(18):7085–7089. doi: 10.1073/pnas.87.18.7085

Intimate association of Thy-1 and the T-cell antigen receptor with the CD45 tyrosine phosphatase.

S Volarević 1, C M Burns 1, J J Sussman 1, J D Ashwell 1
PMCID: PMC54688  PMID: 1976251

Abstract

Immunoprecipitation of Thy-1 from Triton X-100 detergent lysates of surface-iodinated and chemically cross-linked T cells precipitated at least five major and discrete bands. Four of these bands were identified as Thy-1, CD45 (a transmembrane tyrosine phosphatase), a major histocompatibility complex-encoded class I molecule, and beta 2-microglobulin. Similar analyses revealed that CD45 was coprecipitated from lysates of cross-linker-treated cells by antibodies to the T-cell antigen receptor (TCR). The same pattern of coprecipitated bands was observed when digitonin was used to lyse untreated cells. Immunoprecipitation of Thy-1 or the TCR from lysates of cross-linked T cells precipitated CD45 tyrosine phosphatase activity. Calculations based upon the amounts of coprecipitated enzymatic activity or TCR zeta chain indicate that a substantial fraction of Thy-1 and TCR complexes can be cross-linked to CD45. These data support a model in which the dependence of Thy-1 signaling on TCR coexpression is due to their common interaction with a tyrosine phosphatase and provide a possible structural basis for the influence of CD45 on TCR-mediated signaling.

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Selected References

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

  1. Altevogt P., Schreck J., Schraven B., Meuer S., Schirrmacher V., Mitsch A. Association of CD2 and T200 (CD45) in mouse T lymphocytes. Int Immunol. 1990;2(4):353–360. doi: 10.1093/intimm/2.4.353. [DOI] [PubMed] [Google Scholar]
  2. Ashwell J. D., Cunningham R. E., Noguchi P. D., Hernandez D. Cell growth cycle block of T cell hybridomas upon activation with antigen. J Exp Med. 1987 Jan 1;165(1):173–194. doi: 10.1084/jem.165.1.173. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Ashwell J. D., Klusner R. D. Genetic and mutational analysis of the T-cell antigen receptor. Annu Rev Immunol. 1990;8:139–167. doi: 10.1146/annurev.iy.08.040190.001035. [DOI] [PubMed] [Google Scholar]
  4. Bourguignon L. Y., Hyman R., Trowbridge I., Singer S. J. Participation of histocompatibility antigens in capping of molecularly independent cell surface components by their specific antibodies. Proc Natl Acad Sci U S A. 1978 May;75(5):2406–2410. doi: 10.1073/pnas.75.5.2406. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Charbonneau H., Tonks N. K., Walsh K. A., Fischer E. H. The leukocyte common antigen (CD45): a putative receptor-linked protein tyrosine phosphatase. Proc Natl Acad Sci U S A. 1988 Oct;85(19):7182–7186. doi: 10.1073/pnas.85.19.7182. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Gunter K. C., Malek T. R., Shevach E. M. T cell-activating properties of an anti-Thy-1 monoclonal antibody. Possible analogy to OKT3/Leu-4. J Exp Med. 1984 Mar 1;159(3):716–730. doi: 10.1084/jem.159.3.716. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Haustein D., Marchalonis J. J., Harris A. W. Immunoglobulin of T lymphoma cells. Biosynthesis, surface representation, and partial characterization. Biochemistry. 1975 May 6;14(9):1826–1834. doi: 10.1021/bi00680a004. [DOI] [PubMed] [Google Scholar]
  8. Hsi E. D., Siegel J. N., Minami Y., Luong E. T., Klausner R. D., Samelson L. E. T cell activation induces rapid tyrosine phosphorylation of a limited number of cellular substrates. J Biol Chem. 1989 Jun 25;264(18):10836–10842. [PubMed] [Google Scholar]
  9. Hyman R., Trowbridge I. Two complementation classes of T200 (Ly-5) glycoprotein-negative mutants. Immunogenetics. 1981 Mar 1;12(5-6):511–523. doi: 10.1007/BF01561692. [DOI] [PubMed] [Google Scholar]
  10. Kiener P. A., Mittler R. S. CD45-protein tyrosine phosphatase cross-linking inhibits T cell receptor CD3-mediated activation in human T cells. J Immunol. 1989 Jul 1;143(1):23–28. [PubMed] [Google Scholar]
  11. Ledbetter J. A., Rose L. M., Spooner C. E., Beatty P. G., Martin P. J., Clark E. A. Antibodies to common leukocyte antigen p220 influence human T cell proliferation by modifying IL 2 receptor expression. J Immunol. 1985 Sep;135(3):1819–1825. [PubMed] [Google Scholar]
  12. Ledbetter J. A., Tonks N. K., Fischer E. H., Clark E. A. CD45 regulates signal transduction and lymphocyte activation by specific association with receptor molecules on T or B cells. Proc Natl Acad Sci U S A. 1988 Nov;85(22):8628–8632. doi: 10.1073/pnas.85.22.8628. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Leo O., Foo M., Sachs D. H., Samelson L. E., Bluestone J. A. Identification of a monoclonal antibody specific for a murine T3 polypeptide. Proc Natl Acad Sci U S A. 1987 Mar;84(5):1374–1378. doi: 10.1073/pnas.84.5.1374. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Low M. G., Kincade P. W. Phosphatidylinositol is the membrane-anchoring domain of the Thy-1 glycoprotein. Nature. 1985 Nov 7;318(6041):62–64. doi: 10.1038/318062a0. [DOI] [PubMed] [Google Scholar]
  15. Lynes M. A., Karl M., DiBiase K., Flaherty L. Supramolecular relationships of membrane antigens on the murine thymocyte. J Immunogenet. 1982 Dec;9(6):475–484. doi: 10.1111/j.1744-313x.1982.tb01009.x. [DOI] [PubMed] [Google Scholar]
  16. Martorell J., Vilella R., Borche L., Rojo I., Vives J. A second signal for T cell mitogenesis provided by monoclonal antibodies CD45 (T200). Eur J Immunol. 1987 Oct;17(10):1447–1451. doi: 10.1002/eji.1830171010. [DOI] [PubMed] [Google Scholar]
  17. Marvel J., Mayer A. CD45R gives immunofluorescence and transduces signals on mouse T cells. Eur J Immunol. 1988 May;18(5):825–828. doi: 10.1002/eji.1830180526. [DOI] [PubMed] [Google Scholar]
  18. Merćep M., Bonifacino J. S., Garcia-Morales P., Samelson L. E., Klausner R. D., Ashwell J. D. T cell CD3-zeta eta heterodimer expression and coupling to phosphoinositide hydrolysis. Science. 1988 Oct 28;242(4878):571–574. doi: 10.1126/science.2845582. [DOI] [PubMed] [Google Scholar]
  19. Mittler R. S., Greenfield R. S., Schacter B. Z., Richard N. F., Hoffmann M. K. Antibodies to the common leukocyte antigen (T200) inhibit an early phase in the activation of resting human B cells. J Immunol. 1987 May 15;138(10):3159–3166. [PubMed] [Google Scholar]
  20. Mustelin T., Coggeshall K. M., Altman A. Rapid activation of the T-cell tyrosine protein kinase pp56lck by the CD45 phosphotyrosine phosphatase. Proc Natl Acad Sci U S A. 1989 Aug;86(16):6302–6306. doi: 10.1073/pnas.86.16.6302. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Ostergaard H. L., Shackelford D. A., Hurley T. R., Johnson P., Hyman R., Sefton B. M., Trowbridge I. S. Expression of CD45 alters phosphorylation of the lck-encoded tyrosine protein kinase in murine lymphoma T-cell lines. Proc Natl Acad Sci U S A. 1989 Nov;86(22):8959–8963. doi: 10.1073/pnas.86.22.8959. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Pingel J. T., Thomas M. L. Evidence that the leukocyte-common antigen is required for antigen-induced T lymphocyte proliferation. Cell. 1989 Sep 22;58(6):1055–1065. doi: 10.1016/0092-8674(89)90504-7. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Samelson L. E., Patel M. D., Weissman A. M., Harford J. B., Klausner R. D. Antigen activation of murine T cells induces tyrosine phosphorylation of a polypeptide associated with the T cell antigen receptor. Cell. 1986 Sep 26;46(7):1083–1090. doi: 10.1016/0092-8674(86)90708-7. [DOI] [PubMed] [Google Scholar]
  24. Sarmiento M., Dialynas D. P., Lancki D. W., Wall K. A., Lorber M. I., Loken M. R., Fitch F. W. Cloned T lymphocytes and monoclonal antibodies as probes for cell surface molecules active in T cell-mediated cytolysis. Immunol Rev. 1982;68:135–169. doi: 10.1111/j.1600-065x.1982.tb01063.x. [DOI] [PubMed] [Google Scholar]
  25. Schrader J. W., Cunningham B. A., Edelman G. M. Functional interactions of viral and histocompatibility antigens at tumor cell surfaces. Proc Natl Acad Sci U S A. 1975 Dec;72(12):5066–5070. doi: 10.1073/pnas.72.12.5066. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Schraven B., Roux M., Hutmacher B., Meuer S. C. Triggering of the alternative pathway of human T cell activation involves members of the T 200 family of glycoproteins. Eur J Immunol. 1989 Feb;19(2):397–403. doi: 10.1002/eji.1830190226. [DOI] [PubMed] [Google Scholar]
  27. Sparks J. W., Brautigan D. L. Specificity of protein phosphotyrosine phosphatases. Comparison with mammalian alkaline phosphatase using polypeptide substrates. J Biol Chem. 1985 Feb 25;260(4):2042–2045. [PubMed] [Google Scholar]
  28. Sparrow R. L., McKenzie I. F. A function for human T200 in natural killer cytolysis. Transplantation. 1983 Aug;36(2):166–171. doi: 10.1097/00007890-198308000-00011. [DOI] [PubMed] [Google Scholar]
  29. Springer T., Galfrè G., Secher D. S., Milstein C. Monoclonal xenogeneic antibodies to murine cell surface antigens: identification of novel leukocyte differentiation antigens. Eur J Immunol. 1978 Aug;8(8):539–551. doi: 10.1002/eji.1830080802. [DOI] [PubMed] [Google Scholar]
  30. Sussman J. J., Bonifacino J. S., Lippincott-Schwartz J., Weissman A. M., Saito T., Klausner R. D., Ashwell J. D. Failure to synthesize the T cell CD3-zeta chain: structure and function of a partial T cell receptor complex. Cell. 1988 Jan 15;52(1):85–95. doi: 10.1016/0092-8674(88)90533-8. [DOI] [PubMed] [Google Scholar]
  31. Sussman J. J., Saito T., Shevach E. M., Germain R. N., Ashwell J. D. Thy-1- and Ly-6-mediated lymphokine production and growth inhibition of a T cell hybridoma require co-expression of the T cell antigen receptor complex. J Immunol. 1988 Apr 15;140(8):2520–2526. [PubMed] [Google Scholar]
  32. Targan S. R., Newman W. Definition of a "trigger" stage in the NK cytolytic reaction sequence by a monoclonal antibody to the glycoprotein T-200. J Immunol. 1983 Sep;131(3):1149–1153. [PubMed] [Google Scholar]
  33. Thomas M. L. The leukocyte common antigen family. Annu Rev Immunol. 1989;7:339–369. doi: 10.1146/annurev.iy.07.040189.002011. [DOI] [PubMed] [Google Scholar]
  34. Tighe H., Clark M., Waldmann H. Blocking of cytotoxic T cell function by monoclonal antibodies against the CD45 antigen (T200/leukocyte-common antigen). Transplantation. 1987 Dec;44(6):818–823. doi: 10.1097/00007890-198712000-00020. [DOI] [PubMed] [Google Scholar]
  35. Tonks N. K., Charbonneau H., Diltz C. D., Fischer E. H., Walsh K. A. Demonstration that the leukocyte common antigen CD45 is a protein tyrosine phosphatase. Biochemistry. 1988 Nov 29;27(24):8695–8701. doi: 10.1021/bi00424a001. [DOI] [PubMed] [Google Scholar]
  36. Trowbridge I. S., Ralph P., Bevan M. J. Differences in the surface proteins of mouse B and T cells. Proc Natl Acad Sci U S A. 1975 Jan;72(1):157–161. doi: 10.1073/pnas.72.1.157. [DOI] [PMC free article] [PubMed] [Google Scholar]

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