title: Interaction of wheat germ agglutinin with human platelets: a model for studying platelet response.
abstract: Binding of wheat germ agglutinin (WGA) to human platelets and their activation were studied. The binding of fluoresceinyl-substituted WGA (F-WGA) was saturable, specific, reversible and cooperative. The apparent association constant (Ka) was 2.4 X 10(6) M-1. Activation of platelets was measured by change in light transmission (LT) (aggregation) and by assays of 14C-serotonin and beta-thromboglobulin (secretion). The maximum platelet activation was obtained with 15 micrograms/ml WGA. At this concentration only 17% of all available binding sites were occupied. Increasing the WGA concentration diminished the change in LT and the secretion from alpha granules but not from dense bodies. The addition of EDTA partially reduced the platelet aggregation without any effect on the secretion. Remaining LT change was insensitive to all metabolic inhibitors tested (CP/CPK, arginine, indomethacin, PGE1, chlorpromazine, colchicine and cytochalasin B). The secretion measured in the presence of EDTA was only reduced by preincubation with PGE1 and CPZ. Succinyl-WGA which only binds glucoconjugates containing GlcNAc did not induce any platelet activation, whereas limulin, which binds to glucoconjugates containing NeuAc did induce platelet agglutination and secretion. These results indicate that: (1) the platelet aggregation and secretion induced by WGA occur with only partial occupancy of the membrane binding sites by the lectin; (2) the platelet LT change is due in part to the agglutinating properties of the lectin bound to the platelet surface, and in another part to the aggregation reaction mediated by a platelet constituent secreted from the storage sites in response to the binding of the lectin.
mesh_terms: D000527:Alprostadil; D001120:Arginine / Q000494:pharmacology; D001792:Blood Platelets / Q000187:drug effects / Q000378:metabolism*; D002746:Chlorpromazine / Q000494:pharmacology; D003078:Colchicine / Q000494:pharmacology; D003402:Creatine Kinase / Q000494:pharmacology; D003571:Cytochalasin B / Q000494:pharmacology; D004492:Edetic Acid / Q000494:pharmacology; D006801:Humans; D066298:In Vitro Techniques; D007213:Indomethacin / Q000494:pharmacology; D037102:Lectins / Q000494:pharmacology*; D010725:Phosphocreatine / Q000494:pharmacology; D010974:Platelet Aggregation / Q000187:drug effects*; D011458:Prostaglandins E / Q000494:pharmacology; D014909:Wheat Germ Agglutinins
chemical_list: D037102:Lectins; D011458:Prostaglandins E; D014909:Wheat Germ Agglutinins; D010725:Phosphocreatine; D003571:Cytochalasin B; D001120:Arginine; D004492:Edetic Acid; D003402:Creatine Kinase; D000527:Alprostadil; D003078:Colchicine; D002746:Chlorpromazine; D007213:Indomethacin