Human Neutrophil Elastase Mudulates Platelet Function by Limited Proteolysis of Membrane Glycoproteins

Author(s): Levin, R., Garry, K.

Journal/Book: J Ckin Invest. 1985; 75: 657-666.

Abstract: During blond coagulation human polymorphonuclear leukocytes release elastase in amounts that can exceed 100 nmol/liter. We therefore studied the effect of elastase on platelet structure and function. Physiologic concentrations of elastase specifically inhibited thrombin-induced platelet aggregation and ristocetin induced agglutination of ·washed platelets in a time- and dosedependent manner. This was associated with a decrease in the number of high affinity thrombin binding sites on the platelet surface (analysis by "Ligand" program) from 31 per platelet to 12 per platelet (P < 0.05). As analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, treatment of 3H-labeled platelets with elastase resulted in a decrease in the percent glycoprotein at 130,000-150,000 M, = and an increase in the percent protein at M, = 102,000. The supernatant from elastase-treated platelets contained a M, = 88,000 glycoprotein not found in the supernatant from untreated platelets. Immunoprecipitation studies with monoclonal antiglycoprotein lb demonstrated that treatment of ·whole platelets with physiologic concentrations of elastase resulted in proteolytic cleavage of glycoprotein lb. Elastase treatment of glycoprotein immunoisolated with monoclonal antiglycoprotein lb antibody resulted in formation of a glycopeptide with the same electrophoretic mobility as the M, = 102,000 membrane-related glycopeptide. In contrast, analysis by Western blot technique using antiglycoprotein llb and Illa antibodies demonstrated that elastase did not degrade glycoproteins llb or Illa.We conclude that elastase inhibition of thrombin-induced platelet stimulation is accompanied by (a) a reduction in the number of thrombin binding sites per platelet and (b) proteolysis of glycoprotein lb.

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