Tyrosine kinases regulate the cytoskeletal attachment of integrin alpha IIb beta 3 (platelet glycoprotein IIb/IIIa) and the cellular retraction of fibrin polymers.
Integrins promote cell-substratum and cell-cell adhesion by acting as transmembrane linker molecules between extracellular adhesion proteins and the actin-rich cytoskeleton. The integrin alpha IIb beta 3 (platelet glycoprotein IIb/IIIa) is essential for platelet spreading, aggregation, fibrin clot retraction, and for the transduction of extracellular signals. We examined the effect of the specific tyrosine kinase inhibitor herbimycin A on integrin and cytoskeletal-mediated events in thrombin-stimulated platelets. Incubation of washed platelets for 24 h with herbimycin A (5 microM) abolished the thrombin-stimulated cytoskeletal enzyme activity of pp60c-src in parallel with a reduction in the tyrosine phosphorylation of multiple platelet proteins, as assessed with anti-phosphotyrosine immunoblots. However, thrombin-induced activation of protein kinase C and the production of thromboxane A2 were not altered by herbimycin A. Despite the absence of cytoskeletal pp60c-src enzyme activity, platelet shape change, aggregation, and serotonin release were unaltered following platelet stimulation with thrombin (0.05-1.0 unit/ml). Herbimycin A-treated platelets also demonstrated normal platelet aggregation in response to collagen (5 micrograms/ml), ionophore A23187 (2 microM), and ADP/adrenaline (10 microM each). However, the ability of herbimycin A-treated platelets to retract fibrin gels was significantly reduced. This defect in clot retraction was associated with reduced incorporation of integrin alpha IIb beta 3 into the cytoskeletal fraction of thrombin-aggregated platelets. Our studies suggest that tyrosine kinases in platelets regulate the cytoskeletal attachment of alpha IIb beta 3, as an essential process for the transmission of cellular contractile forces to fibrin polymers.
Schoenwaelder
S M
SM
Department of Medicine, Monash Medical School, Box Hill Hospital, Victoria, Australia.
Jackson
S P
SP
Yuan
Y
Y
Teasdale
M S
MS
Salem
H H
HH
Mitchell
C A
CA
D016227:Benzoquinones; D001665:Binding Sites; D001704:Biopolymers / Q000378:metabolism; D001792:Blood Platelets / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure; D001798:Blood Proteins / Q000378:metabolism; D003599:Cytoskeleton / Q000378:metabolism*; D005337:Fibrin / Q000378:metabolism*; D006801:Humans; D047029:Lactams, Macrocyclic; D008855:Microscopy, Electron, Scanning; D010766:Phosphorylation; D010974:Platelet Aggregation / Q000187:drug effects; D010980:Platelet Membrane Glycoproteins / Q000378:metabolism*; D011505:Protein-Tyrosine Kinases / Q000502:physiology*; D011809:Quinones / Q000494:pharmacology; D017828:Rifabutin / Q000031:analogs & derivatives; D012701:Serotonin / Q000378:metabolism; D013917:Thrombin / Q000494:pharmacology
D016227:Benzoquinones; D001704:Biopolymers; D001798:Blood Proteins; D047029:Lactams, Macrocyclic; D010980:Platelet Membrane Glycoproteins; D011809:Quinones; D017828:Rifabutin; D012701:Serotonin; C020754:herbimycin; D005337:Fibrin; D011505:Protein-Tyrosine Kinases; D013917:Thrombin