Activation of stimulus-specific serine esterases (proteases) in the initiation of platelet secretion. I. Demonstration with organophosphorus inhibitors.
The effect of organophosphorus inhibitors of serine esterases (proteases) on secretion from washed rabbit platelets was examined. Five noncytotoxic stimuli were employed: collagen, thrombin, heterologous anti-platelet antibody (in the absence of complement), rabbit C3 bound to zymosan, and platelet activating factor derived from antigen-stimulated, IgE-sensitized rabbit basophils. Diisoprophyl phosphofluoridate, three series of p-nitrophenyl ethyl phosphonates, and a series of cyclohexyl phenylalkylphosphonofluridates were all found to be inhibitory to the platelet secretion. These are irreversible inhibitors of serine proteases but in this system were only inhibitory if added to the platelets concurrently with the stimuli. Pretreatment of either the platelets or the stimuli with the inhibitors followed by washing, was without effect on the subsequent reaction. This suggested the involvement of stimulus-activatable serine proteases in the secretory process. The concept was supported by finding that nonphosphorylating phosphonates or hydrolyzed phosphonates or phosphonofluoridates were without inhibitory action. The effect of a series of phosphonates or phosphonoflouridates in inhibiting each stimulus exhibited a unique activity-structure profile. The demonstration of such unique profiles with four series of inhibitors for each of the five stimuli was interpreted as demonstrating that a specific activatable serine protease was involved in the platelet secretory response to each stimulus.
D000818:Animals; D000906:Antibodies; D001792:Blood Platelets / Q000378:metabolism*; D003094:Collagen / Q000494:pharmacology; D003176:Complement C3; D004950:Esterases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D007531:Isoflurophate / Q000494:pharmacology; D007700:Kinetics; D063065:Organophosphonates / Q000494:pharmacology; D011817:Rabbits; D012636:Secretory Rate / Q000187:drug effects; D012701:Serotonin / Q000378:metabolism*; D013917:Thrombin / Q000037:antagonists & inhibitors / Q000494:pharmacology; D015054:Zymosan / Q000494:pharmacology
D000906:Antibodies; D003176:Complement C3; D063065:Organophosphonates; D007531:Isoflurophate; D012701:Serotonin; D003094:Collagen; D015054:Zymosan; D004950:Esterases; D013917:Thrombin