Detergent-resistant cytoskeleton of the surface-activated platelet differs from the suspension-activated platelet cytoskeleton.
This study contrasts the protein composition of the detergent-resistant cytoskeleton of platelets fully spread on glass with the cytoskeletal composition of resting platelets and platelets aggregated in suspension with thrombin. Complete Triton X-100-insoluble cytoskeletons were isolated from spread, resting, and suspension-activated platelets in the presence of protease inhibitors, solubilized in sodium dodecyl sulfate/EDTA and analyzed on reduced, one-dimensional polyacrylamide gels. The protein composition of the cytoskeletons differed both qualitatively and quantitatively. Most notable were more extensive incorporation of total protein, talin, and vinculin into the cytoskeleton of spread platelets than the cytoskeleton of suspension-activated platelets. Varying the concentration and time of exposure to thrombin during suspension activation did not mimic the cytoskeletal changes of surface activation. Scanning electron microscopy, measurement of lipid phosphorus content, and varying the duration of Triton extraction did not show incomplete solubilization or nonspecific trapping of constituents in the spread platelet cytoskeleton. Proteolysis of talin was minimal in suspension-activated platelets and in platelets spread for 50 minutes. The differences in the detergent-resistant cytoskeletons of surface- and suspension-activated platelets indicate significant divergence in the physiologies of platelet spreading on surfaces and platelet activation in suspension.
Smith
C M
CM
Department of Pediatrics, University of Minnesota Health Sciences Center, Minneapolis 55455.
Burris
S M
SM
Rao
G H
GH
White
J G
JG
D000328:Adult; D001792:Blood Platelets / Q000502:physiology* / Q000648:ultrastructure*; D015153:Blotting, Western; D002458:Cell Fractionation; D003598:Cytoskeletal Proteins / Q000097:blood* / Q000302:isolation & purification; D003599:Cytoskeleton / Q000648:ultrastructure*; D003902:Detergents; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D017830:Octoxynol; D015539:Platelet Activation; D010973:Platelet Adhesiveness; D011092:Polyethylene Glycols; D012995:Solubility; D014461:Ultracentrifugation
D003598:Cytoskeletal Proteins; D003902:Detergents; D011092:Polyethylene Glycols; D017830:Octoxynol