Receptor-dependent and -independent protein phosphorylation in platelets of spontaneously hypertensive rats.
This study investigates the role of protein kinase C and of myosin light chain kinase in mediating platelet hyperresponsiveness in spontaneously hypertensive rats (SHR). For this purpose, 32P-labeled washed platelets of both SHR and normotensive controls Wistar-Kyoto (WKY) were challenged either with a receptor-mediated agonist (thrombin) or with direct activators of myosin light chain kinase and protein kinase C. Such enzymatic activities were assessed by measuring changes in 32P-labeling of their respective target proteins, namely myosin light chain (20 KDa) and the 47 KDa protein. In resting platelets, the patterns of protein phosphorylation were similar between SHR and WKY, suggesting that the two cell types were in a comparable quiescent status. By contrast, in both dose-response and time-course studies, thrombin promoted a significantly greater phosphorylation of the 20- and 47 KDa proteins in platelets of SHR compared with that for WKY. Sensitivity of myosin light chain kinase to the calcium ionophore A23187 and of protein kinase C to both phorbol ester and dioctanoylglycerol was apparently not different between the two cell types. The data indicate that the exaggerated thrombin-induced protein phosphorylation observed for platelets of SHR is not linked to alterations in protein kinase C and/or myosin light chain kinase per se. These results therefore suggest that platelet hyperresponsiveness in SHR is likely to be related, at least in part, to abnormalities in receptor-mediated transmembrane signalling.
Koutouzov
S
S
INSERM U7/CNRS UA 318, Department of Pharmacology, Hospital Necker, Paris, France.
Limon
I
I
Marche
P
P
D000818:Animals; D001792:Blood Platelets / Q000187:drug effects / Q000378:metabolism*; D006973:Hypertension / Q000097:blood*; D008297:Male; D009219:Myosin-Light-Chain Kinase / Q000097:blood; D010750:Phosphoproteins / Q000097:blood*; D011493:Protein Kinase C / Q000097:blood; D051381:Rats; D011918:Rats, Inbred SHR; D011921:Rats, Inbred WKY; D011956:Receptors, Cell Surface / Q000187:drug effects / Q000502:physiology; D013917:Thrombin / Q000494:pharmacology
D010750:Phosphoproteins; D011956:Receptors, Cell Surface; D011493:Protein Kinase C; D009219:Myosin-Light-Chain Kinase; D013917:Thrombin