Inhibition by ajoene of protein tyrosine phosphatase activity in human platelets.
The effects of ajoene (a potent antithrombotic agent obtained from garlic) on the tyrosine phosphorylation status of human platelet proteins were investigated by immunoblotting-based experiments using an anti-phosphotyrosine antibody. Incubation of platelets with ajoene enhanced the phosphorylation of at least four proteins (estimated MWs 76, 80, 84 and 120 kDa), both in resting platelets and in platelets subsequently stimulated with thrombin (0.1 U/ml). This effect was both dose- and incubation-time-dependent. High concentrations of ajoene (50 microM) or long periods of incubation (10 min) led to nonselective 'hyperphosphorylation' of numerous proteins. The effects of ajoene on protein tyrosine phosphatase (PTP) activity in platelet lysates were also investigated, PTP activity was inhibited when platelets were incubated with ajoene before lysis, but not when ajoene was added to lysates of platelets which had not been pre-exposed to ajoene.
Villar
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Departamento de Farmacoloxía, Facultade de Farmacia, Universidade de Santiago de Compostela, Spain. ffvillar@usc.es
Alvariño
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Flores
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D001792:Blood Platelets / Q000187:drug effects* / Q000201:enzymology*; D001798:Blood Proteins / Q000737:chemistry / Q000378:metabolism; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D004220:Disulfides / Q000494:pharmacology*; D005340:Fibrinogen / Q000378:metabolism; D006801:Humans; D066298:In Vitro Techniques; D008560:Membrane Fluidity / Q000187:drug effects; D008970:Molecular Weight; D010766:Phosphorylation; D010936:Plant Extracts / Q000494:pharmacology*; D010974:Platelet Aggregation / Q000187:drug effects; D010975:Platelet Aggregation Inhibitors / Q000494:pharmacology*; D019039:Platelet Glycoprotein GPIIb-IIIa Complex / Q000187:drug effects / Q000378:metabolism; D017027:Protein Tyrosine Phosphatases / Q000037:antagonists & inhibitors* / Q000097:blood*; D013454:Sulfoxides; D013917:Thrombin / Q000494:pharmacology
D001798:Blood Proteins; D004220:Disulfides; D010936:Plant Extracts; D010975:Platelet Aggregation Inhibitors; D019039:Platelet Glycoprotein GPIIb-IIIa Complex; D013454:Sulfoxides; D005340:Fibrinogen; C048980:ajoene; D017027:Protein Tyrosine Phosphatases; D013917:Thrombin