[Increased phosphorylations of proteins involved in the expression of the physiologic response of platelets in SHR rats].
In primary hypertension, phospholipase C (PLC) is hypersensitive in several target tissues (platelets, vascular smooth muscle cells, aortic fibroblasts). Protein kinase C (PKC) and myosin light chain kinase (MLCK), which are physiologically activated by PLC-triggered second messengers (diacylglycerol and Ca2+ ions, respectively), phosphorylate specific proteins closely involved in the cell functional responses. In this study, we have examined and compared between platelets of spontaneously hypertensive rats (SHR) and their normotensive controls Wistar-Kyoto (WKY), the patterns of protein phosphorylation obtained either with the receptor-mediated agonist thrombin (i.e. which acts via PLC) or with direct activators of the protein kinases, PKC and MLCK. Activation by thrombin of 32P-prelabeled platelets induced incorporation of radioactivity into two proteins, P20 (myosin light chain) and P47. The curves obtained when platelets were challenged with either increasing doses of thrombin (0.025-0.3 U/ml) for 20 sec or with a low dose of the agent (0.1 U/ml) for up to 1 min, revealed that phosphorylation of the target proteins of PKC (P47) and of MLCK (P20) were significantly enhanced in platelets of SHR compared to WKY. In contrast, direct activation of PKC by phorbol ester and of MLCK by the calcium ionophore A23187 evoked the selective phosphorylation of the respective target proteins, P47 and P20, to a similar extent in platelets of SHR and WKY. Taken together, these results demonstrate that a physiological agonist (thrombin) induces an enhanced phosphorylation of intracellular proteins in platelets of SHR.(ABSTRACT TRUNCATED AT 250 WORDS)
Koutouzov
S
S
Départment de pharmacologie, INSERM U 7, hôpital Necker, Paris, France.
Limon
I
I
Girard
A
A
Marche
P
P
D000818:Animals; D001792:Blood Platelets / Q000201:enzymology / Q000378:metabolism*; D001798:Blood Proteins / Q000378:metabolism*; D004789:Enzyme Activation; D006973:Hypertension / Q000503:physiopathology; D009219:Myosin-Light-Chain Kinase / Q000378:metabolism; D010761:Phosphorus Radioisotopes; D010766:Phosphorylation; D011494:Protein Kinases / Q000378:metabolism*; D051381:Rats; D011918:Rats, Inbred SHR; D011921:Rats, Inbred WKY; D013917:Thrombin / Q000378:metabolism; D010738:Type C Phospholipases / Q000378:metabolism
D001798:Blood Proteins; D010761:Phosphorus Radioisotopes; D011494:Protein Kinases; D009219:Myosin-Light-Chain Kinase; D010738:Type C Phospholipases; D013917:Thrombin
Augmentation des phosphorylations de protéines impliquées dans l'expression des réponses physiologiques plaquettaires chez le rat SHR.