Interaction of pp60c-src, phospholipase C, inositol-lipid, and diacyglycerol kinases with the cytoskeletons of thrombin-stimulated platelets.
Phosphatidylinositol (PtdIns)-4- and -3-kinases, PtdIns(4)P-5-kinase, diacylglycerol (DAG) kinase, and PtdIns-phospholipase C were all detected in cytoskeletons of resting human platelets. The total cytoskeletal enzyme activities were greatly increased upon thrombin stimulation of the intact cells. Those reached a maximum after a 60-s stimulation for PtdIns(4)P-5-kinase and phospholipase C, while the other kinases appeared to be slightly delayed. Specific activities were stimulated from about 4-fold (PtdIns-3-kinase) to about 6-fold (PtdIns-4-kinase). Thrombin treatment also promoted a co-extraction of pp60c-src with the cytoskeletons and its disappearance from the Triton X-100 soluble fraction. These results suggest that stimulation of platelets by thrombin causes the association of enzymes responsible for lipid phosphorylation and hydrolysis with the cytoskeletons. This could occur at cytoskeleton anchoring points to the membranes.
Grondin
P
P
Institut National de la Santé et de la Recherche Médicale, Unité 326, Hôpital Purpan, Toulouse, France.
Plantavid
M
M
Sultan
C
C
Breton
M
M
Mauco
G
G
Chap
H
H
D019870:1-Phosphatidylinositol 4-Kinase; D001792:Blood Platelets / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism*; D015153:Blotting, Western; D002851:Chromatography, High Pressure Liquid; D003599:Cytoskeleton / Q000201:enzymology / Q000378:metabolism*; D019852:Diacylglycerol Kinase; D006801:Humans; D007700:Kinetics; D019869:Phosphatidylinositol 3-Kinases; D010766:Phosphorylation; D010770:Phosphotransferases / Q000378:metabolism*; D015539:Platelet Activation; D016392:Proto-Oncogene Proteins pp60(c-src) / Q000378:metabolism*; D013917:Thrombin / Q000494:pharmacology; D010738:Type C Phospholipases / Q000378:metabolism*
D010770:Phosphotransferases; D019869:Phosphatidylinositol 3-Kinases; D019852:Diacylglycerol Kinase; D019870:1-Phosphatidylinositol 4-Kinase; D016392:Proto-Oncogene Proteins pp60(c-src); D010738:Type C Phospholipases; D013917:Thrombin