Skip to main content
NCBI home page
Biochemical Journal logoLink to Biochemical Journal
. 1995 May 15;308(Pt 1):347–352. doi: 10.1042/bj3080347

Activation and tyrosine phosphorylation of p72syk as well as calcium mobilization after hydrogen peroxide stimulation in peripheral blood lymphocytes.

S Qin 1, T Inazu 1, H Yamamura 1
PMCID: PMC1136883  PMID: 7538757

Abstract

To determine the regulatory role of p72syk in lymphocyte activation, peripheral blood lymphocytes were treated with 10 mM hydrogen peroxide. Hydrogen peroxide induced a rapid elevation of p72syk activity (4-6-fold) and a dramatic increase in tyrosine phosphorylation of a number of cellular proteins, including phospholipase C gamma 1 (PLC gamma 1) and p72syk. Monoclonal antibodies to PLC gamma 1 co-precipitated p72syk from hydrogen peroxide-stimulated cell lysates, but not from unstimulated cell lysates. Furthermore, we observed a rise in intracellular Ca2+, corresponding to the combination of extracellular Ca2+ influx and the release from intracellular Ca2+ stores. Extracellular Ca2+ influx was necessary for the sustenance of p72syk activity, but not for the initiation of p72syk activation induced by hydrogen peroxide. Taken together, these data suggested that one possible role of p72syk was to activate PLC gamma 1, at least in part through tyrosine phosphorylation, and then to trigger calcium mobilization in pig peripheral blood lymphocytes in response to hydrogen peroxide stimulation.

Full text

PDF
347

Images in this article

348Figure 1
on p.348
349Figure 2
on p.349
349Figure 3
on p.349
350Figure 4
on p.350
350Figure 5
on p.350
350Figure 6
on p.350

Selected References

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

  1. Agarwal A., Salem P., Robbins K. C. Involvement of p72syk, a protein-tyrosine kinase, in Fc gamma receptor signaling. J Biol Chem. 1993 Jul 25;268(21):15900–15905. [PubMed] [Google Scholar]
  2. Bourgoin S., Grinstein S. Peroxides of vanadate induce activation of phospholipase D in HL-60 cells. Role of tyrosine phosphorylation. J Biol Chem. 1992 Jun 15;267(17):11908–11916. [PubMed] [Google Scholar]
  3. Carter R. H., Park D. J., Rhee S. G., Fearon D. T. Tyrosine phosphorylation of phospholipase C induced by membrane immunoglobulin in B lymphocytes. Proc Natl Acad Sci U S A. 1991 Apr 1;88(7):2745–2749. doi: 10.1073/pnas.88.7.2745. [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. Heffetz D., Bushkin I., Dror R., Zick Y. The insulinomimetic agents H2O2 and vanadate stimulate protein tyrosine phosphorylation in intact cells. J Biol Chem. 1990 Feb 15;265(5):2896–2902. [PubMed] [Google Scholar]
  6. Heffetz D., Zick Y. H2O2 potentiates phosphorylation of novel putative substrates for the insulin receptor kinase in intact Fao cells. J Biol Chem. 1989 Jun 15;264(17):10126–10132. [PubMed] [Google Scholar]
  7. Inazu T., Taniguchi T., Yanagi S., Yamamura H. Protein-tyrosine phosphorylation and aggregation of intact human platelets by vanadate with H2O2. Biochem Biophys Res Commun. 1990 Jul 16;170(1):259–263. doi: 10.1016/0006-291x(90)91268-w. [DOI] [PubMed] [Google Scholar]
  8. Kim J. W., Sim S. S., Kim U. H., Nishibe S., Wahl M. I., Carpenter G., Rhee S. G. Tyrosine residues in bovine phospholipase C-gamma phosphorylated by the epidermal growth factor receptor in vitro. J Biol Chem. 1990 Mar 5;265(7):3940–3943. [PubMed] [Google Scholar]
  9. Kolanus W., Romeo C., Seed B. T cell activation by clustered tyrosine kinases. Cell. 1993 Jul 16;74(1):171–183. doi: 10.1016/0092-8674(93)90304-9. [DOI] [PubMed] [Google Scholar]
  10. Koshio O., Akanuma Y., Kasuga M. Hydrogen peroxide stimulates tyrosine phosphorylation of the insulin receptor and its tyrosine kinase activity in intact cells. Biochem J. 1988 Feb 15;250(1):95–101. doi: 10.1042/bj2500095. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Liao F., Shin H. S., Rhee S. G. Tyrosine phosphorylation of phospholipase C-gamma 1 induced by cross-linking of the high-affinity or low-affinity Fc receptor for IgG in U937 cells. Proc Natl Acad Sci U S A. 1992 Apr 15;89(8):3659–3663. doi: 10.1073/pnas.89.8.3659. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Margolis B., Rhee S. G., Felder S., Mervic M., Lyall R., Levitzki A., Ullrich A., Zilberstein A., Schlessinger J. EGF induces tyrosine phosphorylation of phospholipase C-II: a potential mechanism for EGF receptor signaling. Cell. 1989 Jun 30;57(7):1101–1107. doi: 10.1016/0092-8674(89)90047-0. [DOI] [PubMed] [Google Scholar]
  13. Meisenhelder J., Suh P. G., Rhee S. G., Hunter T. Phospholipase C-gamma is a substrate for the PDGF and EGF receptor protein-tyrosine kinases in vivo and in vitro. Cell. 1989 Jun 30;57(7):1109–1122. doi: 10.1016/0092-8674(89)90048-2. [DOI] [PubMed] [Google Scholar]
  14. Nakanishi O., Shibasaki F., Hidaka M., Homma Y., Takenawa T. Phospholipase C-gamma 1 associates with viral and cellular src kinases. J Biol Chem. 1993 May 25;268(15):10754–10759. [PubMed] [Google Scholar]
  15. Nishibe S., Wahl M. I., Hernández-Sotomayor S. M., Tonks N. K., Rhee S. G., Carpenter G. Increase of the catalytic activity of phospholipase C-gamma 1 by tyrosine phosphorylation. Science. 1990 Nov 30;250(4985):1253–1256. doi: 10.1126/science.1700866. [DOI] [PubMed] [Google Scholar]
  16. Nishizuka Y. The molecular heterogeneity of protein kinase C and its implications for cellular regulation. Nature. 1988 Aug 25;334(6184):661–665. doi: 10.1038/334661a0. [DOI] [PubMed] [Google Scholar]
  17. O'Shea J. J., McVicar D. W., Bailey T. L., Burns C., Smyth M. J. Activation of human peripheral blood T lymphocytes by pharmacological induction of protein-tyrosine phosphorylation. Proc Natl Acad Sci U S A. 1992 Nov 1;89(21):10306–10310. doi: 10.1073/pnas.89.21.10306. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Park D. J., Rho H. W., Rhee S. G. CD3 stimulation causes phosphorylation of phospholipase C-gamma 1 on serine and tyrosine residues in a human T-cell line. Proc Natl Acad Sci U S A. 1991 Jun 15;88(12):5453–5456. doi: 10.1073/pnas.88.12.5453. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Pumiglia K. M., Lau L. F., Huang C. K., Burroughs S., Feinstein M. B. Activation of signal transduction in platelets by the tyrosine phosphatase inhibitor pervanadate (vanadyl hydroperoxide). Biochem J. 1992 Sep 1;286(Pt 2):441–449. doi: 10.1042/bj2860441. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Qin S., Inazu T., Yang C., Sada K., Taniguchi T., Yamamura H. Interleukin 2 mediates p72syk activation in peripheral blood lymphocytes. FEBS Lett. 1994 May 30;345(2-3):233–236. doi: 10.1016/0014-5793(94)00450-1. [DOI] [PubMed] [Google Scholar]
  21. Schieven G. L., Kirihara J. M., Burg D. L., Geahlen R. L., Ledbetter J. A. p72syk tyrosine kinase is activated by oxidizing conditions that induce lymphocyte tyrosine phosphorylation and Ca2+ signals. J Biol Chem. 1993 Aug 5;268(22):16688–16692. [PubMed] [Google Scholar]
  22. Schieven G. L., Kirihara J. M., Myers D. E., Ledbetter J. A., Uckun F. M. Reactive oxygen intermediates activate NF-kappa B in a tyrosine kinase-dependent mechanism and in combination with vanadate activate the p56lck and p59fyn tyrosine kinases in human lymphocytes. Blood. 1993 Aug 15;82(4):1212–1220. [PubMed] [Google Scholar]
  23. Secrist J. P., Karnitz L., Abraham R. T. T-cell antigen receptor ligation induces tyrosine phosphorylation of phospholipase C-gamma 1. J Biol Chem. 1991 Jul 5;266(19):12135–12139. [PubMed] [Google Scholar]
  24. Shen Z., Lin C. T., Unkeless J. C. Correlations among tyrosine phosphorylation of Shc, p72syk, PLC-gamma 1, and [Ca2+]i flux in Fc gamma RIIA signaling. J Immunol. 1994 Mar 15;152(6):3017–3023. [PubMed] [Google Scholar]
  25. Takata M., Sabe H., Hata A., Inazu T., Homma Y., Nukada T., Yamamura H., Kurosaki T. Tyrosine kinases Lyn and Syk regulate B cell receptor-coupled Ca2+ mobilization through distinct pathways. EMBO J. 1994 Mar 15;13(6):1341–1349. doi: 10.1002/j.1460-2075.1994.tb06387.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Takeuchi F., Taniguchi T., Maeda H., Fujii C., Takeuchi N., Yamamura H. The lectin concanavalin A stimulates a protein-tyrosine kinase p72syk in peripheral blood lymphocytes. Biochem Biophys Res Commun. 1993 Jul 15;194(1):91–96. doi: 10.1006/bbrc.1993.1789. [DOI] [PubMed] [Google Scholar]
  27. Taniguchi T., Kitagawa H., Yasue S., Yanagi S., Sakai K., Asahi M., Ohta S., Takeuchi F., Nakamura S., Yamamura H. Protein-tyrosine kinase p72syk is activated by thrombin and is negatively regulated through Ca2+ mobilization in platelets. J Biol Chem. 1993 Feb 5;268(4):2277–2279. [PubMed] [Google Scholar]
  28. Wahl M. I., Nishibe S., Suh P. G., Rhee S. G., Carpenter G. Epidermal growth factor stimulates tyrosine phosphorylation of phospholipase C-II independently of receptor internalization and extracellular calcium. Proc Natl Acad Sci U S A. 1989 Mar;86(5):1568–1572. doi: 10.1073/pnas.86.5.1568. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Weber J. R., Bell G. M., Han M. Y., Pawson T., Imboden J. B. Association of the tyrosine kinase LCK with phospholipase C-gamma 1 after stimulation of the T cell antigen receptor. J Exp Med. 1992 Aug 1;176(2):373–379. doi: 10.1084/jem.176.2.373. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Weiss A., Koretzky G., Schatzman R. C., Kadlecek T. Functional activation of the T-cell antigen receptor induces tyrosine phosphorylation of phospholipase C-gamma 1. Proc Natl Acad Sci U S A. 1991 Jul 1;88(13):5484–5488. doi: 10.1073/pnas.88.13.5484. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Zhao Y., Sudol M., Hanafusa H., Krueger J. Increased tyrosine kinase activity of c-Src during calcium-induced keratinocyte differentiation. Proc Natl Acad Sci U S A. 1992 Sep 1;89(17):8298–8302. doi: 10.1073/pnas.89.17.8298. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Zhao Y., Uyttendaele H., Krueger J. G., Sudol M., Hanafusa H. Inactivation of c-Yes tyrosine kinase by elevation of intracellular calcium levels. Mol Cell Biol. 1993 Dec;13(12):7507–7514. doi: 10.1128/mcb.13.12.7507. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Zick Y., Sagi-Eisenberg R. A combination of H2O2 and vanadate concomitantly stimulates protein tyrosine phosphorylation and polyphosphoinositide breakdown in different cell lines. Biochemistry. 1990 Nov 6;29(44):10240–10245. doi: 10.1021/bi00496a013. [DOI] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES