EFFECT OF TUMOR-PROMOTING PHORBOL ESTER, THROMBIN AND VASOPRESSIN ON TRANSLOCATION OF 3 DISTINCT PROTEIN-KINASE-C ISOFORMS IN HUMAN PLATELETS AND REGULATION BY CALCIUM

被引:68
作者
CRABOS, M
FABBRO, D
STABEL, S
ERNE, P
机构
[1] KANTONSSPITAL,DIV CARDIOL,CH-6000 LUZERN 16,SWITZERLAND
[2] UNIV HOSP BASEL,CH-4031 BASEL,SWITZERLAND
[3] CIBA GEIGY AG,DEPT PHARMACEUT RES ONCOL VIROL,CH-4002 BASEL,SWITZERLAND
[4] MAX PLANCK GESELL,MAX DELBRUCK LAB,W-5000 COLOGNE 30,GERMANY
关键词
D O I
10.1042/bj2880891
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Protein kinase C (PKC) acts in synergy with Ca2+ mobilization for the activation of platelets. Three different PKC subtypes that specifically react with antibodies to alpha- beta- and zeta-PKC have been detected in human platelets. We have compared the subcellular redistribution of these isoforms in platelets after exposure to the tumour-promoting phorbol ester phorbol 12-myristate 13-acetate (PMA) and to two physiological agonists, thrombin and vasopressin. In the presence of PMA, beta-PKC is most rapidly translocated to membranes, followed by zeta-PKC and alpha-PKC [membrane contents of 39+/-6, 31+/-4 and 24+/-4% (means+/-S.E.M.) respectively after 2 min incubation]. In contrast, both thrombin and vasopressin induced a biphasic translocation of PKC isoforms. For both agonists, the first phase of translocation occurred within 1 min and was identical for the three isoforms. However, during the second phase, the translocation of zeta-PKC by thrombin and vasopressin differed [membrane contents (mean+/-S.E.M.) of 24+/-3 and 46+/-4 % respectively after 10 min]. These results suggest a differential activation of zeta-PKC by vasopressin and thrombin. PMA-induced translocation of alpha-PKC was decreased from 278+/-27 to 198+/-24 (mean+/-S.E.M., P = 0.02; percentage increase over control value) in the presence of 1 mM-EDTA, whereas chelation of intracellular Ca2+ by Quin2-AM does not influence this response. These results suggest that the PMA-induced translocation of alpha-PKC depends on the presence of 1 mm concentration of extracellular Ca2+. In addition, the chelation of either extracellular or intracellular Ca2+ inhibited both vasopressin- and thrombin-induced translocation of all three isoforms, suggesting that Ca2+ is an important requirement for the translocation of alpha-, beta- and zeta-PKC by physiological agonists. In conclusion, the translocation of PKC varies between different isoforms and between different agonists.
引用
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页码:891 / 896
页数:6
相关论文
共 42 条
  • [1] ISOLATION AND CHARACTERIZATION OF PKC-L, A NEW MEMBER OF THE PROTEIN-KINASE C-RELATED GENE FAMILY SPECIFICALLY EXPRESSED IN LUNG, SKIN, AND HEART
    BACHER, N
    ZISMAN, Y
    BERENT, E
    LIVNEH, E
    [J]. MOLECULAR AND CELLULAR BIOLOGY, 1991, 11 (01) : 126 - 133
  • [2] IMMUNOLOGICAL QUANTITATION OF PHOSPHOLIPID/CA-2+-DEPENDENT PROTEIN-KINASE OF HUMAN MAMMARY-CARCINOMA CELLS - INVERSE RELATIONSHIP TO ESTROGEN-RECEPTORS
    BORNER, C
    WYSS, R
    REGAZZI, R
    EPPENBERGER, U
    FABBRO, D
    [J]. INTERNATIONAL JOURNAL OF CANCER, 1987, 40 (03) : 344 - 348
  • [3] EFFECTOR-DEPENDENT CONFORMATIONAL-CHANGES IN PROTEIN KINASE-C-GAMMA THROUGH EPITOPE MAPPING WITH INHIBITORY MONOCLONAL-ANTIBODIES
    CAZAUBON, S
    WEBSTER, C
    CAMOIN, L
    STROSBERG, AD
    PARKER, P
    [J]. EUROPEAN JOURNAL OF BIOCHEMISTRY, 1990, 194 (03): : 799 - 804
  • [4] DIFFERENT TRANSLOCATION OF 3 DISTINCT PKC ISOFORMS WITH TUMOR-PROMOTING PHORBOL ESTER IN HUMAN PLATELETS
    CRABOS, M
    IMBER, R
    WOODTLI, T
    FABBRO, D
    ERNE, P
    [J]. BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1991, 178 (03) : 878 - 883
  • [5] ALKYL-LINKED DIGLYCERIDES INHIBIT PROTEIN KINASE-C ACTIVATION BY DIACYLGLYCEROLS
    DANIEL, LW
    SMALL, GW
    SCHMITT, JD
    MARASCO, CJ
    ISHAQ, K
    PIANTADOSI, C
    [J]. BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1988, 151 (01) : 291 - 297
  • [6] DECOURCELLES DD, 1987, THROMB HAEMOSTASIS, V58, P927
  • [7] CALCIUM TRANSIENTS IN HUMAN-PLATELETS MONITORED BY AEQUORIN, FURA-2 AND QUIN-2 - EFFECTS OF PROTEIN-KINASE-C ACTIVATION AND INHIBITION
    ERNE, P
    SCHACHTER, M
    FABBRO, D
    MILES, CMM
    SEVER, PS
    [J]. BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1987, 145 (01) : 66 - 72
  • [8] TRANSLOCATION OF PROTEIN-KINASE-C IS NOT REQUIRED TO INHIBIT THE ANTIGEN-INDUCED INCREASE OF CYTOSOLIC CALCIUM IN A MAST-CELL LINE
    ERNE, P
    MAZUREK, N
    BORNER, C
    CONSCIENCE, JF
    EPPENBERGER, U
    FABBRO, D
    [J]. BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1987, 143 (01) : 252 - 259
  • [9] FORD DA, 1989, J BIOL CHEM, V264, P13818
  • [10] PHORBOL ESTER INDUCES DIFFERENTIAL MEMBRANE-ASSOCIATION OF PROTEIN KINASE-C SUBSPECIES IN HUMAN-PLATELETS
    FOURNIER, A
    HARDY, SJ
    CLARK, KJ
    MURRAY, AW
    [J]. BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1989, 161 (02) : 556 - 561