Neutrophil elastase potentiates cathepsin G-induced platelet activation.
We have previously demonstrated that human neutrophil cathepsin G is a strong platelet agonist that binds to a specific receptor. This work describes the effect of neutrophil elastase on cathepsin G-induced platelet responses. While platelets were not activated by high concentrations of neutrophil elastase by itself, elastase enhanced aggregation, secretion and calcium mobilization induced by low concentrations of cathepsin G. Platelet aggregation and secretion were potentiated in a concentration-dependent manner by neutrophil elastase with maximal responses observable at 200 nM. Enhancement was observed when elastase was preincubated with platelets for time intervals of 10-60 s prior to addition of a low concentration of cathepsin G and required catalytically-active elastase since phenylmethanesulphonyl fluoride-inhibited enzyme failed to potentiate cell activation. Neutrophil elastase potentiation of platelet responses induced by low concentrations of cathepsin G was markedly inhibited by creatine phosphate/creatine phosphokinase and/or indomethacin, indicating that the synergism between elastase and cathepsin G required the participation of ADP and thromboxane A2. On the other hand, platelet responses were not attenuated by the PAF antagonist BN 52021, signifying that PAF-acether did not play a role in elastase potentiation. At higher concentrations porcine pancreatic elastase exhibits similar effects to neutrophil elastase, demonstrating that the effect of elastase was not unique to the neutrophil protease. While neutrophil elastase failed to alter the ability of cathepsin G to hydrolyze a synthetic chromogenic substrate, preincubation of platelets with elastase increased the apparent affinity of cathepsin G binding to platelets.(ABSTRACT TRUNCATED AT 250 WORDS)
Selak
M A
MA
Department of Pharmacology, Temple University Medical School, Philadelphia, Pennsylvania.
D000244:Adenosine Diphosphate / Q000097:blood; D000595:Amino Acid Sequence; D056649:Cathepsin G; D002403:Cathepsins / Q000008:administration & dosage* / Q000378:metabolism; D000242:Cyclic AMP / Q000097:blood; D004357:Drug Synergism; D006801:Humans; D066298:In Vitro Techniques; D008969:Molecular Sequence Data; D009504:Neutrophils / Q000201:enzymology; D009842:Oligopeptides / Q000737:chemistry; D010196:Pancreatic Elastase / Q000008:administration & dosage* / Q000097:blood; D010972:Platelet Activating Factor / Q000378:metabolism; D015539:Platelet Activation / Q000187:drug effects* / Q000502:physiology; D012697:Serine Endopeptidases; D013379:Substrate Specificity
D009842:Oligopeptides; D010972:Platelet Activating Factor; D000244:Adenosine Diphosphate; D000242:Cyclic AMP; D002403:Cathepsins; D012697:Serine Endopeptidases; C534094:CTSG protein, human; D056649:Cathepsin G; D010196:Pancreatic Elastase