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. 1989 Sep 1;262(2):449–456. doi: 10.1042/bj2620449

Complexing of the CD-3 subunit by a monoclonal antibody activates a microtubule-associated protein 2 (MAP-2) serine kinase in Jurkat cells.

C Hanekom 1, A Nel 1, C Gittinger 1, A Rheeder 1, G Landreth 1
PMCID: PMC1133288  PMID: 2552997

Abstract

Treatment of Jurkat T-cells with anti-CD-3 monoclonal antibodies resulted in the rapid and transient activation of a serine kinase which utilized the microtubule-associated protein, MAP-2, as a substrate in vitro. The kinase was also activated on treatment of Jurkat cells with phytohaemagglutinin, but with a different time course. The activation of the MAP-2 kinase by anti-CD-3 antibodies was dose-dependent, with maximal activity observed at concentrations of greater than 500 ng/ml. Normal human E-rosette-positive T-cells also exhibited induction of MAP-2 kinase activity during anti-CD-3 treatment. The enzyme was optimally active in the presence of 2 mM-Mn2+; lower levels of activity were observed with Mg2+, even at concentrations up to 20 mM. The kinase was partially purified by passage over DE-52 Sephacel with the activity eluting as a single peak at 0.25 M-NaCl. The molecular mass was estimated to be 45 kDa by gel filtration. The activation of the MAP-2 kinase was probably due to phosphorylation of this enzyme as treatment with alkaline phosphatase diminished its activity. These data demonstrate that the stimulation of T-cells through the CD-3 complex results in the activation of a novel serine kinase which may be critically involved in signal transduction in these cells.

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Selected References

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

  1. Cheng H. C., Kemp B. E., Pearson R. B., Smith A. J., Misconi L., Van Patten S. M., Walsh D. A. A potent synthetic peptide inhibitor of the cAMP-dependent protein kinase. J Biol Chem. 1986 Jan 25;261(3):989–992. [PubMed] [Google Scholar]
  2. Cheung R. K., Grinstein S., Gelfand E. W. Permissive role of calcium in the inhibition of T cell mitogenesis by calmodulin antagonists. J Immunol. 1983 Nov;131(5):2291–2295. [PubMed] [Google Scholar]
  3. Cheung W. Y. Calmodulin: an overview. Fed Proc. 1982 May;41(7):2253–2257. [PubMed] [Google Scholar]
  4. Cochet C., Gill G. N., Meisenhelder J., Cooper J. A., Hunter T. C-kinase phosphorylates the epidermal growth factor receptor and reduces its epidermal growth factor-stimulated tyrosine protein kinase activity. J Biol Chem. 1984 Feb 25;259(4):2553–2558. [PubMed] [Google Scholar]
  5. 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]
  6. Czech M. P., Klarlund J. K., Yagaloff K. A., Bradford A. P., Lewis R. E. Insulin receptor signaling. Activation of multiple serine kinases. J Biol Chem. 1988 Aug 15;263(23):11017–11020. [PubMed] [Google Scholar]
  7. Ek B., Westermark B., Wasteson A., Heldin C. H. Stimulation of tyrosine-specific phosphorylation by platelet-derived growth factor. Nature. 1982 Feb 4;295(5848):419–420. doi: 10.1038/295419a0. [DOI] [PubMed] [Google Scholar]
  8. Fukunaga K., Yamamoto H., Matsui K., Higashi K., Miyamoto E. Purification and characterization of a Ca2+- and calmodulin-dependent protein kinase from rat brain. J Neurochem. 1982 Dec;39(6):1607–1617. doi: 10.1111/j.1471-4159.1982.tb07994.x. [DOI] [PubMed] [Google Scholar]
  9. Gilman A. G. G proteins and dual control of adenylate cyclase. Cell. 1984 Mar;36(3):577–579. doi: 10.1016/0092-8674(84)90336-2. [DOI] [PubMed] [Google Scholar]
  10. Hidaka H., Inagaki M., Kawamoto S., Sasaki Y. Isoquinolinesulfonamides, novel and potent inhibitors of cyclic nucleotide dependent protein kinase and protein kinase C. Biochemistry. 1984 Oct 9;23(21):5036–5041. doi: 10.1021/bi00316a032. [DOI] [PubMed] [Google Scholar]
  11. Hunter T. A thousand and one protein kinases. Cell. 1987 Sep 11;50(6):823–829. doi: 10.1016/0092-8674(87)90509-5. [DOI] [PubMed] [Google Scholar]
  12. 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]
  13. Iwashita S., Fox C. F. Epidermal growth factor and potent phorbol tumor promoters induce epidermal growth factor receptor phosphorylation in a similar but distinctively different manner in human epidermoid carcinoma A431 cells. J Biol Chem. 1984 Feb 25;259(4):2559–2567. [PubMed] [Google Scholar]
  14. Kadowaki T., Fujita-Yamaguchi Y., Nishida E., Takaku F., Akiyama T., Kathuria S., Akanuma Y., Kasuga M. Phosphorylation of tubulin and microtubule-associated proteins by the purified insulin receptor kinase. J Biol Chem. 1985 Apr 10;260(7):4016–4020. [PubMed] [Google Scholar]
  15. Kasuga M., Fujita-Yamaguchi Y., Blithe D. L., Kahn C. R. Tyrosine-specific protein kinase activity is associated with the purified insulin receptor. Proc Natl Acad Sci U S A. 1983 Apr;80(8):2137–2141. doi: 10.1073/pnas.80.8.2137. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Kim H., Binder L. I., Rosenbaum J. L. The periodic association of MAP2 with brain microtubules in vitro. J Cell Biol. 1979 Feb;80(2):266–276. doi: 10.1083/jcb.80.2.266. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Klemm D. J., Elias L. A distinctive phospholipid-stimulated protein kinase of normal and malignant murine hemopoietic cells. J Biol Chem. 1987 Jun 5;262(16):7580–7585. [PubMed] [Google Scholar]
  18. Kuo J. F., Schatzman R. C., Turner R. S., Mazzei G. J. Phospholipid-sensitive Ca2+-dependent protein kinase: a major protein phosphorylation system. Mol Cell Endocrinol. 1984 May;35(2-3):65–73. doi: 10.1016/0303-7207(84)90001-7. [DOI] [PubMed] [Google Scholar]
  19. 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]
  20. Landreth G. E., Rieser G. D. Nerve growth factor- and epidermal growth factor-stimulated phosphorylation of a PC12 cytoskeletally associated protein in situ. J Cell Biol. 1985 Mar;100(3):677–683. doi: 10.1083/jcb.100.3.677. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Landreth G. E., Williams L. K. Nerve growth factor stimulates the phosphorylation of a 250 kDa cytoskeletal protein in cell-free extracts of PC12 cells. Neurochem Res. 1987 Oct;12(10):943–950. doi: 10.1007/BF00966317. [DOI] [PubMed] [Google Scholar]
  22. Manger B., Weiss A., Weyand C., Goronzy J., Stobo J. D. T cell activation: differences in the signals required for IL 2 production by nonactivated and activated T cells. J Immunol. 1985 Dec;135(6):3669–3673. [PubMed] [Google Scholar]
  23. Nel A. E., Bouic P., Lattanze G. R., Stevenson H. C., Miller P., Dirienzo W., Stefanini G. F., Galbraith R. M. Reaction of T lymphocytes with anti-T3 induces translocation of C-kinase activity to the membrane and specific substrate phosphorylation. J Immunol. 1987 May 15;138(10):3519–3524. [PubMed] [Google Scholar]
  24. Nel A. E., Vandenplas M., Wooten M. M., Cooper R., Vandenplas S., Rheeder A., Daniels J. Cholera toxin partially inhibits the T-cell response to phytohaemagglutinin through the ADP-ribosylation of a 45 kDa membrane protein. Biochem J. 1988 Dec 1;256(2):383–390. doi: 10.1042/bj2560383. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Nel A. E., Wooten M. W., Galbraith R. M. Molecular signaling mechanisms in T-lymphocyte activation pathways: a review and future prospects. Clin Immunol Immunopathol. 1987 Aug;44(2):167–186. doi: 10.1016/0090-1229(87)90064-x. [DOI] [PubMed] [Google Scholar]
  26. Nishida E., Hoshi M., Miyata Y., Sakai H., Kadowaki T., Kasuga M., Saijo S., Ogawara H., Akiyama T. Tyrosine phosphorylation by the epidermal growth factor receptor kinase induces functional alterations in microtubule-associated protein 2. J Biol Chem. 1987 Nov 25;262(33):16200–16204. [PubMed] [Google Scholar]
  27. Nishizuka Y. The role of protein kinase C in cell surface signal transduction and tumour promotion. Nature. 1984 Apr 19;308(5961):693–698. doi: 10.1038/308693a0. [DOI] [PubMed] [Google Scholar]
  28. O'Flynn K., Russul-Saib M., Ando I., Wallace D. L., Beverley P. C., Boylston A. W., Linch D. C. Different pathways of human T-cell activation revealed by PHA-P and PHA-M. Immunology. 1986 Jan;57(1):55–60. [PMC free article] [PubMed] [Google Scholar]
  29. Olmsted J. B. Microtubule-associated proteins. Annu Rev Cell Biol. 1986;2:421–457. doi: 10.1146/annurev.cb.02.110186.002225. [DOI] [PubMed] [Google Scholar]
  30. Pellegrino M. A., Ferrone S., Dierich M. P., Reisfeld R. A. Enhancement of sheep red blood cell human lymphocyte rosette formation by the sulfhydryl compound 2-amino ethylisothiouronium bromide. Clin Immunol Immunopathol. 1975 Jan;3(3):324–333. doi: 10.1016/0090-1229(75)90019-7. [DOI] [PubMed] [Google Scholar]
  31. Pettey C. L., Alarcon B., Malin R., Weinberg K., Terhorst C. T3-p28 is a protein associated with the delta and epsilon chains of the T cell receptor-T3 antigen complex during biosynthesis. J Biol Chem. 1987 Apr 5;262(10):4854–4859. [PubMed] [Google Scholar]
  32. Ray L. B., Sturgill T. W. Characterization of insulin-stimulated microtubule-associated protein kinase. Rapid isolation and stabilization of a novel serine/threonine kinase from 3T3-L1 cells. J Biol Chem. 1988 Sep 5;263(25):12721–12727. [PubMed] [Google Scholar]
  33. Ray L. B., Sturgill T. W. Insulin-stimulated microtubule-associated protein kinase is phosphorylated on tyrosine and threonine in vivo. Proc Natl Acad Sci U S A. 1988 Jun;85(11):3753–3757. doi: 10.1073/pnas.85.11.3753. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Ray L. B., Sturgill T. W. Rapid stimulation by insulin of a serine/threonine kinase in 3T3-L1 adipocytes that phosphorylates microtubule-associated protein 2 in vitro. Proc Natl Acad Sci U S A. 1987 Mar;84(6):1502–1506. doi: 10.1073/pnas.84.6.1502. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Sibley D. R., Strasser R. H., Caron M. G., Lefkowitz R. J. Homologous desensitization of adenylate cyclase is associated with phosphorylation of the beta-adrenergic receptor. J Biol Chem. 1985 Apr 10;260(7):3883–3886. [PubMed] [Google Scholar]
  36. Sturgill T. W., Ray L. B., Erikson E., Maller J. L. Insulin-stimulated MAP-2 kinase phosphorylates and activates ribosomal protein S6 kinase II. Nature. 1988 Aug 25;334(6184):715–718. doi: 10.1038/334715a0. [DOI] [PubMed] [Google Scholar]
  37. Theurkauf W. E., Vallee R. B. Extensive cAMP-dependent and cAMP-independent phosphorylation of microtubule-associated protein 2. J Biol Chem. 1983 Jun 25;258(12):7883–7886. [PubMed] [Google Scholar]
  38. Theurkauf W. E., Vallee R. B. Molecular characterization of the cAMP-dependent protein kinase bound to microtubule-associated protein 2. J Biol Chem. 1982 Mar 25;257(6):3284–3290. [PubMed] [Google Scholar]
  39. Tsoukas C. D., Landgraf B., Bentin J., Valentine M., Lotz M., Vaughan J. H., Carson D. A. Activation of resting T lymphocytes by anti-CD3 (T3) antibodies in the absence of monocytes. J Immunol. 1985 Sep;135(3):1719–1723. [PubMed] [Google Scholar]
  40. Tsuyama S., Bramblett G. T., Huang K. P., Flavin M. Calcium/phospholipid-dependent kinase recognizes sites in microtubule-associated protein 2 which are phosphorylated in living brain and are not accessible to other kinases. J Biol Chem. 1986 Mar 25;261(9):4110–4116. [PubMed] [Google Scholar]
  41. Ushiro H., Cohen S. Identification of phosphotyrosine as a product of epidermal growth factor-activated protein kinase in A-431 cell membranes. J Biol Chem. 1980 Sep 25;255(18):8363–8365. [PubMed] [Google Scholar]
  42. Valentine M. A., Tsoukas C. D., Rhodes G., Vaughan J. H., Carson D. A. Phytohemagglutinin binds to the 20-kDa molecule of the T3 complex. Eur J Immunol. 1985 Aug;15(8):851–854. doi: 10.1002/eji.1830150821. [DOI] [PubMed] [Google Scholar]
  43. Vallee R. B. A taxol-dependent procedure for the isolation of microtubules and microtubule-associated proteins (MAPs). J Cell Biol. 1982 Feb;92(2):435–442. doi: 10.1083/jcb.92.2.435. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Weiss A., Imboden J. B. Cell surface molecules and early events involved in human T lymphocyte activation. Adv Immunol. 1987;41:1–38. doi: 10.1016/s0065-2776(08)60029-2. [DOI] [PubMed] [Google Scholar]
  45. Yamauchi T., Fujisawa H. Phosphorylation of microtubule-associated protein 2 by calmodulin-dependent protein kinase (Kinase II) which occurs only in the brain tissues. Biochem Biophys Res Commun. 1982 Dec 15;109(3):975–981. doi: 10.1016/0006-291x(82)92035-6. [DOI] [PubMed] [Google Scholar]
  46. Yu K. T., Khalaf N., Czech M. P. Insulin stimulates a novel Mn2+-dependent cytosolic serine kinase in rat adipocytes. J Biol Chem. 1987 Dec 5;262(34):16677–16685. [PubMed] [Google Scholar]

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