scholarly journals A protein kinase C pseudosubstrate peptide inhibits phosphorylation of the CD3 antigen in streptolysin-O-permeabilized human T lymphocytes

1989 ◽  
Vol 260 (3) ◽  
pp. 893-901 ◽  
Author(s):  
D R Alexander ◽  
J M Hexham ◽  
S C Lucas ◽  
J D Graves ◽  
D A Cantrell ◽  
...  
Keyword(s):  
T Cells ◽  
Protein Kinase C ◽  
Protein Kinase ◽  
T Lymphocytes ◽  
Gamma Chain ◽  
Permeabilized Cells ◽  
Human T Lymphocytes ◽  
Kinase C ◽  
Streptolysin O ◽  
Subsequent Addition

Activation of human T lymphocytes leads to the phosphorylation of the CD3-antigen gamma polypeptide. We have investigated a possible role for protein kinase C (PKC) in mediating this phosphorylation event by using T cells permeabilized with streptolysin-O in the presence of 120 mM-K+ buffers containing Ca2+-EGTA. The gamma-chain was phosphorylated by [gamma-32P]ATP in permeabilized T lymphoblasts in the presence of phorbol 12,13-dibutyrate (Pdbu) or phytohaemagglutinin (PHA). Ca2+ alone in the range 0.5-1.0 microM also induced gamma-chain phosphorylation in some T-lymphoblast preparations; that in Jurkat-6 cells occurred at lower concentrations (50-500 nM). Two experimental approaches were used to investigate the possible involvement of PKC. Firstly, when permeabilization was carried out in buffer lacking free Ca2+, PKC was lost from the cells, and gamma-chain phosphorylation could then no longer be induced on subsequent addition of Pdbu or PHA in 400 nM-Ca2+, or 800 nM-Ca2+ alone, to permeabilized cells. However, when permeabilization was carried out in the presence of these three agents, PKC was translocated to intracellular membranes, and subsequent addition of [gamma-32P]ATP to these cells then resulted in gamma-chain phosphorylation. In the second approach, induction of gamma-chain phosphorylation by Pdbu, 1-oleoyl-2-acetylglycerol, 1,2-diolein, PHA or Ca2+ alone was effectively blocked by permeabilizing T cells in the presence of a PKC pseudosubstrate peptide (50 microM). Pseudosubstrate concentrations in the range 7-20 microM inhibited gamma-chain phosphorylation by 50%. In contrast, addition of four other ‘irrelevant’ basic peptides (50 microM) did not result in detectable inhibition, and 50 microM-pseudosubstrate did not inhibit the phosphorylation of 17 other polypeptides isolated from permeabilized T cells. These data suggest that Pdbu-, 1,2-diacylglycerol-, PHA- and Ca2+-induced phosphorylation of the CD3-antigen gamma chain in permeabilized T cells is mediated by PKC.

Activation-dependent phosphorylation of endogeneous protein kinase c substrates in quiescent human T lymphocytes

Immunobiology ◽  
1988 ◽  
Vol 176 (4-5) ◽  
pp. 465-478 ◽  
Author(s):  
Bengt Friedrich ◽  
Kristina Noreus ◽  
Doreen A. Cantrell ◽  
Martin Gullberg
Keyword(s):  
Protein Kinase C ◽  
Protein Kinase ◽  
T Lymphocytes ◽  
Human T Lymphocytes ◽  
Kinase C

scholarly journals Chromatin-Associated Protein Kinase C-θ Regulates an Inducible Gene Expression Program and MicroRNAs in Human T Lymphocytes

2011 ◽  
Vol 41 (6) ◽  
pp. 704-719 ◽  
Author(s):  
Elissa L. Sutcliffe ◽  
Karen L. Bunting ◽  
Yi Qing He ◽  
Jasmine Li ◽  
Chansavath Phetsouphanh ◽  
...  
Keyword(s):  
Gene Expression ◽  
Protein Kinase C ◽  
Protein Kinase ◽  
T Lymphocytes ◽  
Gene Expression Program ◽  
Inducible Gene Expression ◽  
Human T Lymphocytes ◽  
Kinase C ◽  
Expression Program ◽  
Inducible Gene

scholarly journals Mechanisms for Induction of L-Selectin Loss from T Lymphocytes by a Cryptococcal Polysaccharide, Glucuronoxylomannan

1999 ◽  
Vol 67 (1) ◽  
pp. 220-229 ◽  
Author(s):  
Zhao Ming Dong ◽  
Lydgia Jackson ◽  
Juneann W. Murphy
Keyword(s):  
T Cells ◽  
Protein Kinase C ◽  
Protein Kinase ◽  
T Lymphocytes ◽  
Tyrosine Kinases ◽  
Jurkat Cells ◽  
Protein Tyrosine Kinases ◽  
Protein Tyrosine ◽  
Herbimycin A ◽  
Kinase C

ABSTRACT Disseminated cryptococcosis is accompanied by cryptococcal polysaccharides in the serum and the lack of cellular infiltrates in infected tissues. Cryptococcal polysaccharides given intravenously to mice inhibit the influx of T lymphocytes into the sites of cell-mediated immune response. The focus here was to determine whether cryptococcal polysaccharides modulate the expression of molecules, such as L-selectin, that are important in extravasation of T cells. Cryptococcal glucuronoxylomannan (GXM), but not galactoxylomannan or mannoprotein, was found to cause loss of L-selectin from freshly isolated human T cells of both CD4 and CD8 subsets and from Jurkat cells. With the signaling-pathway inhibitors staurosporine (which inhibits protein kinase C) and herbimycin A (which inhibits protein tyrosine kinases), we showed that GXM or the cryptococcal culture filtrate antigen CneF directly induces L-selectin loss from CD4+ and CD8+ T cells via a herbimycin A-sensitive pathway(s) presumably involving one or more protein tyrosine kinases but not via a pathway involving protein kinase C. Loss of L-selectin from the T cells before the T cells have a chance to bind to L-selectin ligands on endothelial cells would be expected to prevent T-cell migration into inflamed tissues and/or lymph organs.

scholarly journals Isolation of protein kinase C subspecies from a preparation of human T lymphocytes

FEBS Letters ◽  
1988 ◽  
Vol 234 (2) ◽  
pp. 387-391 ◽  
Author(s):  
Mark S. Shearman ◽  
Nicola Berry ◽  
Tomiichiro Oda ◽  
Katsuhiko Ase ◽  
Ushio Kikkawa ◽  
...  
Keyword(s):  
Protein Kinase C ◽  
Protein Kinase ◽  
T Lymphocytes ◽  
Human T Lymphocytes ◽  
Kinase C

Comparison of interleukin 2 and 12-O-tetradecanoyl-phorbol 13-acetate as signals for protein kinase C activation in purified human T lymphocytes

1988 ◽  
Vol 18 (12) ◽  
pp. 2029-2036 ◽  
Author(s):  
Yann Mahé ◽  
Jean-Pierre Piau ◽  
Naomi Wakasugi ◽  
Thomas Tursz ◽  
Gerard Gacon ◽  
...  
Keyword(s):  
Protein Kinase C ◽  
Protein Kinase ◽  
T Lymphocytes ◽  
Interleukin 2 ◽  
Human T Lymphocytes ◽  
Kinase C ◽  
Protein Kinase C Activation

Proliferative responses induced by the activation of protein kinase C during the development of human T lymphocytes

1991 ◽  
Vol 21 (1) ◽  
pp. 115-121 ◽  
Author(s):  
Africa González-Fernández ◽  
Fernando Diaz-Espada ◽  
Miguel Kreisler ◽  
Francisco Gambón Deza
Keyword(s):  
Protein Kinase C ◽  
Protein Kinase ◽  
T Lymphocytes ◽  
Human T Lymphocytes ◽  
Kinase C

Activators of protein kinase C and 5-azacytidine induce IL-2 receptor expression on human T lymphocytes

1985 ◽  
Vol 27 (3) ◽  
pp. 267-276 ◽  
Author(s):  
Joel M. Depper ◽  
Warren J. Leonard ◽  
Cynthia L. Drogula ◽  
Martin Krönke ◽  
Thomas A. Waldmann ◽  
...  
Keyword(s):  
Protein Kinase C ◽  
Protein Kinase ◽  
T Lymphocytes ◽  
Receptor Expression ◽  
Human T Lymphocytes ◽  
Kinase C

scholarly journals A method for measuring protein kinase C activity in permeabilized T lymphocytes by using peptide substrates. Evidence for multiple pathways of kinase activation

1990 ◽  
Vol 268 (2) ◽  
pp. 303-308 ◽  
Author(s):  
D R Alexander ◽  
J D Graves ◽  
S C Lucas ◽  
D A Cantrell ◽  
M J Crumpton
Keyword(s):  
T Cells ◽  
Protein Kinase C ◽  
Protein Kinase ◽  
T Lymphocytes ◽  
Inositol Phosphates ◽  
Assay System ◽  
Peptide Substrates ◽  
Kinase C ◽  
Wide Range ◽  
Pkc Activation

Activation of protein kinase C (PKC) in human T lymphocytes is an immediate consequence of mitogenic signalling via the antigen-receptor complex and CD2 antigen. In order to investigate further the signal-transduction pathways which result in PKC activation, we have established a novel PKC assay system using streptolysin-O-permeabilized T cells. Known peptide substrates of PKC were introduced into permeabilized cells in the presence of [gamma-32P]ATP, 3 mM-Mg2+ and 150 nM free Ca2+. The peptide found to have the lowest background phosphorylation had the sequence Pro-Leu-Ser-Arg-Thr-Leu-Ser-Val-Ala-Ala-Lys-Lys (peptide GS), and the phosphorylation of the peptide was increased up to 6-fold by direct activation of PKC with phorbol 12,13-dibutyrate. Induction of PKC activation with the UCHT1 antibody against the CD3 antigen, or with phytohaemagglutinin (PHA) or guanosine 5′-[gamma-thio]triphosphate (GTP[S]), increased peptide-GS phosphorylation by 2-3 fold. The specificity of PKC action on peptide GS was demonstrated by blocking increases in phosphorylation with a pseudosubstrate peptide PKC inhibitor. PKC activation by this technique could be detected within 1 min of adding external ligand. Dose-response curves revealed that PHA-induced production of inositol phosphates correlated closely with PKC activities, whereas only a partial correlation between these parameters was observed with GTP[S]. Our data are consistent with the presence of more than one G-protein-mediated pathway of PKC regulation in T cells. The quantitative PKC assay system described is both simple and reproducible, and its potential application to a wide range of cell types should prove useful in further investigations of PKC activation mechanisms.

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