EVIDENCE FOR REQUIREMENT OF TYROSINE PHOSPHORYLATION IN ENDOTHELIAL P-2Y-PURINOCEPTOR AND P-2U-PURINOCEPTOR STIMULATION OF PROSTACYCLIN RELEASE

被引:17
作者
BOWDEN, A [1 ]
PATEL, V [1 ]
BROWN, C [1 ]
BOARDER, MR [1 ]
机构
[1] UNIV LEICESTER,DEPT CELL PHYSIOL & PHARMACOL,LEICESTER LE1 9HN,LEICS,ENGLAND
关键词
P-2-PURINOCEPTORS; PROSTACYCLIN; PHOSPHOLIPASE A(2); ENDOTHELIUM; TYROSINE KINASES;
D O I
10.1111/j.1476-5381.1995.tb17208.x
中图分类号
R9 [药学];
学科分类号
1007 ;
摘要
1 The release of prostacylin (PGI(2)) from vascular endothelial cells is stimulated by ATP acting at G protein-coupled P-2-purinoceptors. Here we investigate the hypothesis that tyrosine protein phosphorylations are involved in this response. 2 The use of Western blots with anti-phosphotyrosine antibodies showed that 30 mu M 2MeSATP (selective for P-2Y-purinoceptors), 300 mu M UTP (selective for P-2U-purinoceptors) and 300 mu M ATP (effective at both these purinoceptors), each stimulate the tyrosine phosphorylation of proteins in bovine cultured aortic endothelial cells. Each of these agonists also stimulates 6-keto PGF(1 alpha) accumulation in the medium (an index of PGI(2) release) in these cells in the same period. 3 The tyrosine kinase inhibitor, genistein, inhibits the 6-keto PGF(1 alpha) response with the same concentration-dependency (1-100 mu M) as the tyrosine phosphorylation response. 4 Tyrphostin, a structurally and functionally distinct tyrosine kinase inhibitor, is also a potent inhibitor (0.1-10 mu M) of the 6-keto PGF(1 alpha) response. 5 Neither tyrphostin nor genistein inhibit the phospholipase C response to P-2-purinoceptor stimulation. Furthermore, these inhibitors do not affect the 6-keto PGF(1 alpha) response to ionomycin. 6 These results show that the regulation of vascular endothelial cells by ATP acting at both P-2Y- and P-2U-purinoceptors involves the stimulation of tyrosine phosphorylation, and suggest that this is a necessary event for the purinoceptor-mediated stimulation of PGI(2) production.
引用
收藏
页码:2563 / 2568
页数:6
相关论文
共 31 条
[1]  
AKIYAMA T, 1987, J BIOL CHEM, V262, P5592
[2]   CULTURE OF ARTERIAL ENDOTHELIAL CELLS - CHARACTERIZATION AND GROWTH OF BOVINE AORTIC CELLS [J].
BOOYSE, FM ;
SEDLAK, BJ ;
RAFELSON, ME .
THROMBOSIS ET DIATHESIS HAEMORRHAGICA, 1975, 34 (03) :825-839
[3]   PROTEIN KINASE-C ACTIVATION ALTERS THE SENSITIVITY OF AGONIST-STIMULATED ENDOTHELIAL-CELL PROSTACYCLIN PRODUCTION TO INTRACELLULAR CA-2+ [J].
CARTER, TD ;
HALLAM, TJ ;
PEARSON, JD .
BIOCHEMICAL JOURNAL, 1989, 262 (02) :431-437
[4]   REGULATION OF P-2Y-PURINOCEPTOR-MEDIATED PROSTACYCLIN RELEASE FROM HUMAN-ENDOTHELIAL CELLS BY CYTOPLASMIC CALCIUM-CONCENTRATION [J].
CARTER, TD ;
HALLAM, TJ ;
CUSACK, NJ ;
PEARSON, JD .
BRITISH JOURNAL OF PHARMACOLOGY, 1988, 95 (04) :1181-1190
[5]  
CHEN YH, 1994, J BIOL CHEM, V269, P27372
[6]   RAS-DEPENDENT ACTIVATION OF MAP KINASE PATHWAY MEDIATED BY G-PROTEIN BETA-GAMMA-SUBUNITS [J].
CRESPO, P ;
XU, NZ ;
SIMONDS, WF ;
GUTKIND, JS .
NATURE, 1994, 369 (6479) :418-420
[7]  
DUFF JL, 1994, J BIOL CHEM, V268, P26037
[8]  
FORCE T, 1991, J BIOL CHEM, V266, P10754
[9]   TYRPHOSTINS .1. SYNTHESIS AND BIOLOGICAL-ACTIVITY OF PROTEIN TYROSINE KINASE INHIBITORS [J].
GAZIT, A ;
YAISH, P ;
GILON, C ;
LEVITZKI, A .
JOURNAL OF MEDICINAL CHEMISTRY, 1989, 32 (10) :2344-2352
[10]  
GRANOT Y, 1993, J BIOL CHEM, V268, P9564