Shear-stress-induced von Willebrand factor binding to platelets causes the activation of tyrosine kinase(s).
Pathological arterial blood flow generates fluid shear stresses that directly cause platelet aggregation. The mechanism of shear-induced platelet aggregation is incompletely understood, but involves von Willebrand factor (vWF) binding to platelet glycoprotein (GP) Ib and GP IIb-IIIa, leading to the transmembrane influx of Ca2+ and the activation of protein kinase C. To investigate this further, shear-stress-induced protein tyrosine phosphorylation (PTP) of washed platelets was studied in a cone-plate viscometer. A time- and shear-stress-dependent tyrosine phosphorylation of substrates with approx. M(r) 29,000-31,000, 36,000, 50,000, 58,000, 64,000, 76,000, 85,000 and 105,000 was observed. PTP in response to a threshold shear stress of 0.3 mN/cm2 (30 dyn/cm2) was enhanced in most cases by exogenous purified human vWF, and PTP in response to a pathological shear stress of 0.9 mN/cm2 (90 dyn/cm2) was inhibited in some cases by inhibiting vWF binding to GP Ib or GP IIb-IIIa, or by inhibiting Ca2+ responses with extracellular EGTA. Shear-induced PTP of a substrate of M(r) approximately 31,000 appeared to be independent of GP Ib, and PTP of a substrate(s) of M(r) approximately 29,000 was shear-stress-dependent but independent of extracellular Ca2+. Cytochalasin D, which inhibits GP Ib-cytoskeleton interactions, inhibits the PTP of a substrate of M(r) approximately 76,000. These results suggest that tyrosine phosphorylation may be involved in transmembrane signalling that mediates platelet adhesion and aggregation in response to pathological shear stresses generated at sites of arterial vaso-occlusion.
Razdan
K
K
Veterans' Affairs Medical Center, Houston, TX.
Hellums
J D
JD
Kroll
M H
MH
D001792:Blood Platelets / Q000378:metabolism*; D001809:Blood Viscosity; D015638:Cytochalasin D / Q000494:pharmacology; D004533:Egtazic Acid / Q000494:pharmacology; D004587:Electrophoresis, Agar Gel; D004789:Enzyme Activation; D006801:Humans; D015151:Immunoblotting; D008970:Molecular Weight; D010766:Phosphorylation; D019000:Phosphotyrosine; D010974:Platelet Aggregation / Q000502:physiology; D010980:Platelet Membrane Glycoproteins / Q000378:metabolism*; D011505:Protein-Tyrosine Kinases / Q000378:metabolism*; D013314:Stress, Mechanical; D014443:Tyrosine / Q000031:analogs & derivatives* / Q000378:metabolism; D014841:von Willebrand Factor / Q000378:metabolism*
D010980:Platelet Membrane Glycoproteins; D014841:von Willebrand Factor; D019000:Phosphotyrosine; D015638:Cytochalasin D; D014443:Tyrosine; D004533:Egtazic Acid; D011505:Protein-Tyrosine Kinases