Presence of calmodulin in human platelet cytoskeletons and its concentration change upon activation of platelets.
We found that a small, reproducible amount of calmodulin is present in the cytoskeleton of human platelets. Triton-insoluble materials (cytoskeletons), which were prepared by centrifugation at 1000 X g for 10 min of platelets after lysis by Triton X-100, stimulated cyclic AMP phosphodiesterase activity in the presence of Ca2+ but not in the presence of the calcium chelator, EGTA, or the calmodulin antagonist, trifluoperazine. The activation of the enzyme was also obtained after heating Triton-insoluble materials. An alkaline glycerol polyacrylamide gel electrophoresis of fractions obtained after gel filtration of solubilized Triton residues showed a protein band which had a faster electrophoretic mobility in the absence than in the presence of Ca2+. Upon thrombin activation of platelets, calmodulin in the Triton-insoluble cytoskeletons increased rapidly parallel to actin, actin-binding protein and myosin. With other stimulants such as collagen, epinephrine and ADP, similar results were obtained but with slower association of these proteins with cytoskeletons. However, after treatment with the Ca2+-ionophore A23187, calmodulin, actin and actin-binding protein in Triton residues decreased rapidly, whereas the association of myosin increased. Thus, calmodulin seems to be associated with actin filaments rather than myosin filaments, and may be involved in the generation of contractile force in the cell.
D015105:3',5'-Cyclic-AMP Phosphodiesterases / Q000097:blood; D000244:Adenosine Diphosphate / Q000494:pharmacology; D000251:Adenosine Triphosphatases / Q000097:blood; D001792:Blood Platelets / Q000187:drug effects / Q000502:physiology / Q000648:ultrastructure*; D001798:Blood Proteins / Q000032:analysis; D002147:Calmodulin / Q000097:blood*; D003094:Collagen / Q000494:pharmacology; D003599:Cytoskeleton / Q000502:physiology / Q000648:ultrastructure*; D004789:Enzyme Activation; D004837:Epinephrine / Q000494:pharmacology; D006801:Humans; D007700:Kinetics; D010974:Platelet Aggregation / Q000187:drug effects; D014268:Trifluoperazine / Q000494:pharmacology
D001798:Blood Proteins; D002147:Calmodulin; D014268:Trifluoperazine; D000244:Adenosine Diphosphate; D003094:Collagen; D015105:3',5'-Cyclic-AMP Phosphodiesterases; D000251:Adenosine Triphosphatases; C026049:EDTA-ATPase; D004837:Epinephrine