Skip to main content
NCBI home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1989 May;86(9):3277–3281. doi: 10.1073/pnas.86.9.3277

The CD4 and CD8 antigens are coupled to a protein-tyrosine kinase (p56lck) that phosphorylates the CD3 complex.

E K Barber 1, J D Dasgupta 1, S F Schlossman 1, J M Trevillyan 1, C E Rudd 1
PMCID: PMC287114  PMID: 2470098

Abstract

Many mammalian receptors have been found to regulate cell growth by virtue of a protein-tyrosine kinase domain in their cytoplasmic tail. We recently described an association of the CD4 antigen with a T-cell-specific protein-tyrosine kinase (p56lck; formerly termed pp58lck; EC 2.7.1.112). This interaction represents a potential mechanism by which T-cell growth may be regulated and offers a model by which other members of the src family (products of c-src, c-yes, c-fgr, etc.) may interact with mammalian growth factor receptors. As in the case of the CD4 antigen, the CD8 antigen appears to serve as a receptor for nonpolymorphic regions of products of the major histocompatibility complex and has been implicated in the regulation of T-cell growth. In this study, we reveal that the human CD8 antigen is also associated with the T-cell-specific protein-tyrosine kinase (p56lck). The associated p56lck kinase was detected by use of both in vitro and in vivo labeling regimes using an antiserum to the C terminus of p56lck. Two-dimensional nonequilibrium pH-gradient gel electrophoresis and sodium dodecyl sulfate/polyacrylamide gel electrophoresis demonstrated the similarity of p56lck to the protein-tyrosine kinase associated with the CD4 antigen. The catalytic activity of p56lck was revealed by the autophosphorylation of the 55- to 60-kDa kinase and the occasional labeling of a 35-kDa protein. Last, we demonstrate directly that members of the CD3 complex, including the gamma, delta, and epsilon chains, as well as a putative zeta subunit, can be phosphorylated at tyrosine residues by the CD4/CD8.p56lck complex.

Full text

PDF
3277

Images in this article

3279Image
on p.3279
3279Image
on p.3279
3279Image
on p.3279
3279Image
on p.3279
3279Image
on p.3279
3280Image
on p.3280
3280Image
on p.3280
3280Image
on p.3280
3280Image
on p.3280
3280Image
on p.3280

Selected References

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

  1. Amiot M., Dastot H., Degos L., Dausset J., Bernard A., Boumsell L. HLA class I molecules are associated with CD1a heavy chains on normal human thymus cells. Proc Natl Acad Sci U S A. 1988 Jun;85(12):4451–4454. doi: 10.1073/pnas.85.12.4451. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Amrein K. E., Sefton B. M. Mutation of a site of tyrosine phosphorylation in the lymphocyte-specific tyrosine protein kinase, p56lck, reveals its oncogenic potential in fibroblasts. Proc Natl Acad Sci U S A. 1988 Jun;85(12):4247–4251. doi: 10.1073/pnas.85.12.4247. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Anderson P., Blue M. L., Morimoto C., Schlossman S. F. Cross-linking of T3 (CD3) with T4 (CD4) enhances the proliferation of resting T lymphocytes. J Immunol. 1987 Aug 1;139(3):678–682. [PubMed] [Google Scholar]
  4. Bank I., Chess L. Perturbation of the T4 molecule transmits a negative signal to T cells. J Exp Med. 1985 Oct 1;162(4):1294–1303. doi: 10.1084/jem.162.4.1294. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bishop J. M. Cellular oncogenes and retroviruses. Annu Rev Biochem. 1983;52:301–354. doi: 10.1146/annurev.bi.52.070183.001505. [DOI] [PubMed] [Google Scholar]
  6. Blue M. L., Craig K. A., Anderson P., Branton K. R., Jr, Schlossman S. F. Evidence for specific association between class I major histocompatibility antigens and the CD8 molecules of human suppressor/cytotoxic cells. Cell. 1988 Jul 29;54(3):413–421. doi: 10.1016/0092-8674(88)90204-8. [DOI] [PubMed] [Google Scholar]
  7. Bonifacino J. S., Lippincott-Schwartz J., Chen C., Antusch D., Samelson L. E., Klausner R. D. Association and dissociation of the murine T cell receptor associated protein (TRAP). Early events in the biosynthesis of a multisubunit receptor. J Biol Chem. 1988 Jun 25;263(18):8965–8971. [PubMed] [Google Scholar]
  8. 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]
  9. Cantrell D. A., Davies A. A., Crumpton M. J. Activators of protein kinase C down-regulate and phosphorylate the T3/T-cell antigen receptor complex of human T lymphocytes. Proc Natl Acad Sci U S A. 1985 Dec;82(23):8158–8162. doi: 10.1073/pnas.82.23.8158. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Carlsson J., Drevin H., Axén R. Protein thiolation and reversible protein-protein conjugation. N-Succinimidyl 3-(2-pyridyldithio)propionate, a new heterobifunctional reagent. Biochem J. 1978 Sep 1;173(3):723–737. doi: 10.1042/bj1730723. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. 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]
  12. Cooper J. A., Sefton B. M., Hunter T. Detection and quantification of phosphotyrosine in proteins. Methods Enzymol. 1983;99:387–402. doi: 10.1016/0076-6879(83)99075-4. [DOI] [PubMed] [Google Scholar]
  13. Courtneidge S. A., Smith A. E. Polyoma virus transforming protein associates with the product of the c-src cellular gene. Nature. 1983 Jun 2;303(5916):435–439. doi: 10.1038/303435a0. [DOI] [PubMed] [Google Scholar]
  14. Eichmann K., Jönsson J. I., Falk I., Emmrich F. Effective activation of resting mouse T lymphocytes by cross-linking submitogenic concentrations of the T cell antigen receptor with either Lyt-2 or L3T4. Eur J Immunol. 1987 May;17(5):643–650. doi: 10.1002/eji.1830170510. [DOI] [PubMed] [Google Scholar]
  15. Fleischer B., Schrezenmeier H., Wagner H. Function of the CD4 and CD8 molecules on human cytotoxic T lymphocytes: regulation of T cell triggering. J Immunol. 1986 Mar 1;136(5):1625–1628. [PubMed] [Google Scholar]
  16. Gold D. P., Clevers H., Alarcon B., Dunlap S., Novotny J., Williams A. F., Terhorst C. Evolutionary relationship between the T3 chains of the T-cell receptor complex and the immunoglobulin supergene family. Proc Natl Acad Sci U S A. 1987 Nov;84(21):7649–7653. doi: 10.1073/pnas.84.21.7649. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Hunter T., Cooper J. A. Protein-tyrosine kinases. Annu Rev Biochem. 1985;54:897–930. doi: 10.1146/annurev.bi.54.070185.004341. [DOI] [PubMed] [Google Scholar]
  18. Koga Y., Caccia N., Toyonaga B., Spolski R., Yanagi Y., Yoshikai Y., Mak T. W. A human T cell-specific cDNA clone (YT16) encodes a protein with extensive homology to a family of protein-tyrosine kinases. Eur J Immunol. 1986 Dec;16(12):1643–1646. doi: 10.1002/eji.1830161229. [DOI] [PubMed] [Google Scholar]
  19. Krangel M. S. Endocytosis and recycling of the T3-T cell receptor complex. The role of T3 phosphorylation. J Exp Med. 1987 Apr 1;165(4):1141–1159. doi: 10.1084/jem.165.4.1141. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Krissansen G. W., Owen M. J., Verbi W., Crumpton M. J. Primary structure of the T3 gamma subunit of the T3/T cell antigen receptor complex deduced from cDNA sequences: evolution of the T3 gamma and delta subunits. EMBO J. 1986 Aug;5(8):1799–1808. doi: 10.1002/j.1460-2075.1986.tb04429.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. 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]
  22. Ledbetter J. A., Seaman W. E., Tsu T. T., Herzenberg L. A. Lyt-2 and lyt-3 antigens are on two different polypeptide subunits linked by disulfide bonds. Relationship of subunits to T cell cytolytic activity. J Exp Med. 1981 Jun 1;153(6):1503–1516. doi: 10.1084/jem.153.6.1503. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Ledbetter J. A., Tsu T. T., Clark E. A. Covalent association between human thymus leukemia-like antigens and CD8(Tp32) molecules. J Immunol. 1985 Jun;134(6):4250–4254. [PubMed] [Google Scholar]
  24. Link M. P., Stewart S. J., Warnke R. A., Levy R. Discordance between surface and cytoplasmic expression of the Leu-4 (T3) antigen in thymocytes and in blast cells from childhood T lymphoblastic malignancies. J Clin Invest. 1985 Jul;76(1):248–253. doi: 10.1172/JCI111954. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Littman D. R., Thomas Y., Maddon P. J., Chess L., Axel R. The isolation and sequence of the gene encoding T8: a molecule defining functional classes of T lymphocytes. Cell. 1985 Feb;40(2):237–246. doi: 10.1016/0092-8674(85)90138-2. [DOI] [PubMed] [Google Scholar]
  26. Marrack P., Endres R., Shimonkevitz R., Zlotnik A., Dialynas D., Fitch F., Kappler J. The major histocompatibility complex-restricted antigen receptor on T cells. II. Role of the L3T4 product. J Exp Med. 1983 Oct 1;158(4):1077–1091. doi: 10.1084/jem.158.4.1077. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Marth J. D., Lewis D. B., Wilson C. B., Gearn M. E., Krebs E. G., Perlmutter R. M. Regulation of pp56lck during T-cell activation: functional implications for the src-like protein tyrosine kinases. EMBO J. 1987 Sep;6(9):2727–2734. doi: 10.1002/j.1460-2075.1987.tb02566.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Marth J. D., Peet R., Krebs E. G., Perlmutter R. M. A lymphocyte-specific protein-tyrosine kinase gene is rearranged and overexpressed in the murine T cell lymphoma LSTRA. Cell. 1985 Dec;43(2 Pt 1):393–404. doi: 10.1016/0092-8674(85)90169-2. [DOI] [PubMed] [Google Scholar]
  29. O'Farrell P. Z., Goodman H. M., O'Farrell P. H. High resolution two-dimensional electrophoresis of basic as well as acidic proteins. Cell. 1977 Dec;12(4):1133–1141. doi: 10.1016/0092-8674(77)90176-3. [DOI] [PubMed] [Google Scholar]
  30. Owens T., Fazekas de St Groth B. Participation of L3T4 in T cell activation in the absence of class II major histocompatibility complex antigens. Inhibition by anti-L3T4 antibodies is a function both of epitope density and mode of presentation of anti-receptor antibody. J Immunol. 1987 Apr 15;138(8):2402–2409. [PubMed] [Google Scholar]
  31. Parham P., Bodmer W. F. Monoclonal antibody to a human histocompatibility alloantigen, HLA-A2. Nature. 1978 Nov 23;276(5686):397–399. doi: 10.1038/276397a0. [DOI] [PubMed] [Google Scholar]
  32. Reinherz E. L., Meuer S. C., Schlossman S. F. The human T cell receptor: analysis with cytotoxic T cell clones. Immunol Rev. 1983;74:83–112. doi: 10.1111/j.1600-065x.1983.tb01085.x. [DOI] [PubMed] [Google Scholar]
  33. Rudd C. E., Bodmer J. G., Bodmer W. F., Crumpton M. J. HLA-D region antigen-associated invariant polypeptides as revealed by two-dimensional gel analysis. Glycosylation and structural inter-relationships. J Biol Chem. 1985 Feb 10;260(3):1927–1936. [PubMed] [Google Scholar]
  34. Rudd C. E., Trevillyan J. M., Dasgupta J. D., Wong L. L., Schlossman S. F. The CD4 receptor is complexed in detergent lysates to a protein-tyrosine kinase (pp58) from human T lymphocytes. Proc Natl Acad Sci U S A. 1988 Jul;85(14):5190–5194. doi: 10.1073/pnas.85.14.5190. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Schneider C., Newman R. A., Sutherland D. R., Asser U., Greaves M. F. A one-step purification of membrane proteins using a high efficiency immunomatrix. J Biol Chem. 1982 Sep 25;257(18):10766–10769. [PubMed] [Google Scholar]
  36. Snary D., Goodfellow P., Bodmer W. F., Crumpton M. J. Evidence against a dimeric structure for membrane-bound HLA antigens. Nature. 1975 Nov 20;258(5532):240–242. doi: 10.1038/258240a0. [DOI] [PubMed] [Google Scholar]
  37. Snow P. M., Terhorst C. The T8 antigen is a multimeric complex of two distinct subunits on human thymocytes but consists of homomultimeric forms on peripheral blood T lymphocytes. J Biol Chem. 1983 Dec 10;258(23):14675–14681. [PubMed] [Google Scholar]
  38. Snow P. M., Van de Rijn M., Terhorst C. Association between the human thymic differentiation antigens T6 and T8. Eur J Immunol. 1985 May;15(5):529–532. doi: 10.1002/eji.1830150520. [DOI] [PubMed] [Google Scholar]
  39. Sukhatme V. P., Sizer K. C., Vollmer A. C., Hunkapiller T., Parnes J. R. The T cell differentiation antigen Leu-2/T8 is homologous to immunoglobulin and T cell receptor variable regions. Cell. 1985 Mar;40(3):591–597. doi: 10.1016/0092-8674(85)90207-7. [DOI] [PubMed] [Google Scholar]
  40. Swain S. L. T cell subsets and the recognition of MHC class. Immunol Rev. 1983;74:129–142. doi: 10.1111/j.1600-065x.1983.tb01087.x. [DOI] [PubMed] [Google Scholar]
  41. Swarup G., Dasgupta J. D., Garbers D. L. Tyrosine protein kinase activity of rat spleen and other tissues. J Biol Chem. 1983 Sep 10;258(17):10341–10347. [PubMed] [Google Scholar]
  42. Trevillyan J. M., Canna C., Maley D., Linna T. J., Phillips C. A. Identification of the human T-lymphocyte protein-tyrosine kinase by peptide-specific antibodies. Biochem Biophys Res Commun. 1986 Oct 15;140(1):392–398. doi: 10.1016/0006-291x(86)91103-4. [DOI] [PubMed] [Google Scholar]
  43. Trevillyan J. M., Lin Y., Chen S. J., Phillips C. A., Canna C., Linna T. J. Human T lymphocytes express a protein-tyrosine kinase homologous to p56LSTRA. Biochim Biophys Acta. 1986 Oct 10;888(3):286–295. doi: 10.1016/0167-4889(86)90228-4. [DOI] [PubMed] [Google Scholar]
  44. Veillette A., Bookman M. A., Horak E. M., Bolen J. B. The CD4 and CD8 T cell surface antigens are associated with the internal membrane tyrosine-protein kinase p56lck. Cell. 1988 Oct 21;55(2):301–308. doi: 10.1016/0092-8674(88)90053-0. [DOI] [PubMed] [Google Scholar]
  45. Veillette A., Horak I. D., Horak E. M., Bookman M. A., Bolen J. B. Alterations of the lymphocyte-specific protein tyrosine kinase (p56lck) during T-cell activation. Mol Cell Biol. 1988 Oct;8(10):4353–4361. doi: 10.1128/mcb.8.10.4353. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Voronova A. F., Sefton B. M. Expression of a new tyrosine protein kinase is stimulated by retrovirus promoter insertion. Nature. 1986 Feb 20;319(6055):682–685. doi: 10.1038/319682a0. [DOI] [PubMed] [Google Scholar]
  47. Weiss A., Imboden J., Hardy K., Manger B., Terhorst C., Stobo J. The role of the T3/antigen receptor complex in T-cell activation. Annu Rev Immunol. 1986;4:593–619. doi: 10.1146/annurev.iy.04.040186.003113. [DOI] [PubMed] [Google Scholar]
  48. Weissman A. M., Baniyash M., Hou D., Samelson L. E., Burgess W. H., Klausner R. D. Molecular cloning of the zeta chain of the T cell antigen receptor. Science. 1988 Feb 26;239(4843):1018–1021. doi: 10.1126/science.3278377. [DOI] [PubMed] [Google Scholar]
  49. Weissman A. M., Ross P., Luong E. T., Garcia-Morales P., Jelachich M. L., Biddison W. E., Klausner R. D., Samelson L. E. Tyrosine phosphorylation of the human T cell antigen receptor zeta-chain: activation via CD3 but not CD2. J Immunol. 1988 Nov 15;141(10):3532–3536. [PubMed] [Google Scholar]
  50. White M. F., Maron R., Kahn C. R. Insulin rapidly stimulates tyrosine phosphorylation of a Mr-185,000 protein in intact cells. Nature. 1985 Nov 14;318(6042):183–186. doi: 10.1038/318183a0. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES