Skip to main content
NCBI home page
The Journal of Experimental Medicine logoLink to The Journal of Experimental Medicine
. 1976 Dec 1;144(6):1657–1673. doi: 10.1084/jem.144.6.1657

Activation of stimulus-specific serine esterases (proteases) in the initiation of platelet secretion. I. Demonstration with organophosphorus inhibitors

PMCID: PMC2190490  PMID: 1003107

Abstract

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.

Full Text

The Full Text of this article is available as a PDF (1.3 MB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Ardlie N. G., Packham M. A., Mustard J. F. Adenosine diphosphate-induced platelet aggregation in suspensions of washed rabbit platelets. Br J Haematol. 1970 Jul;19(1):7–17. doi: 10.1111/j.1365-2141.1970.tb01596.x. [DOI] [PubMed] [Google Scholar]
  2. BECKER E. L., FUKUTO T. R., BOONE B., CANHAM D. C., BOGER E. The relationship of enzyme inhibitory activity to the structure of n-alkylphosphonate and phenylalkyl-phosphonate esters. Biochemistry. 1963 Jan-Feb;2:72–76. doi: 10.1021/bi00901a014. [DOI] [PubMed] [Google Scholar]
  3. Becker E. L., Austen K. F. Mechanisms of immunologic injury of rat peritoneal mast cells. I. The effect of phosphonate inhibitors on the homocytotropic antibody-mediated histamine release and the first component of rat complement. J Exp Med. 1966 Sep 1;124(3):379–395. doi: 10.1084/jem.124.3.379. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Becker E. L. The relationship of the structure of phosphonate esters to their ability to inhibit chymotrypsin, trypsin, acetylcholinesterase and C'Ia. Biochim Biophys Acta. 1967 Oct 23;147(2):289–296. doi: 10.1016/0005-2795(67)90407-2. [DOI] [PubMed] [Google Scholar]
  5. Becker E. L., Unanue E. R. The requirement for esterase activation in the anti-immunoglobulin-triggered movement of B lymphocytes. J Immunol. 1976 Jul;117(1):27–32. [PubMed] [Google Scholar]
  6. Benveniste J., Henson P. M., Cochrane C. G. Leukocyte-dependent histamine release from rabbit platelets. The role of IgE, basophils, and a platelet-activating factor. J Exp Med. 1972 Dec 1;136(6):1356–1377. doi: 10.1084/jem.136.6.1356. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Cooper N. R., Becker E. L. Complement associated peptidase activity of guinea pig serum. I. Role of complement components. J Immunol. 1967 Jan;98(1):119–131. [PubMed] [Google Scholar]
  8. Ferluga J., Asherson G. L., Becker E. L. The effect of organophosphorus inhibitors, p-nitrophenol and cytochalasin B on cytotoxic killing of tumour cells by immune spleen cells, and the effect of shaking. Immunology. 1972 Oct;23(4):577–590. [PMC free article] [PubMed] [Google Scholar]
  9. Hafner L. S., Brown J. E., Garrison M. V., Alexander B. H. The synthesis of ethyl p-nitrophenyl alpha-acetoxyalkylphosphonates. J Med Chem. 1965 Sep;8(5):730–731. doi: 10.1021/jm00329a051. [DOI] [PubMed] [Google Scholar]
  10. Henson P. M. Activation and desensitization of platelets by platelet-activating factor (PAF) derived from IgE-sensitized basophils. I. Characteristics of the secretory response. J Exp Med. 1976 Apr 1;143(4):937–952. doi: 10.1084/jem.143.4.937. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Henson P. M. Mechanisms of release of constituents from rabbit platelets by antigen-antibody complexes and complement. I. Lytic and nonlytic reactions. J Immunol. 1970 Aug;105(2):476–489. [PubMed] [Google Scholar]
  12. Henson P. M., Oades Z. G. Activation of platelets by platelet-activating factor (PAF) derived from IgE-sensitized basophils. II. The role of serine proteases, cyclic nucleotides, and contractile elements in PAF-induced secretion. J Exp Med. 1976 Apr 1;143(4):953–968. doi: 10.1084/jem.143.4.953. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Henson P. M. Release of vasoactive amines from rabbit platelets induced by antiplatelet antibody in the presence and absence of complement. J Immunol. 1970 Apr;104(4):924–934. [PubMed] [Google Scholar]
  14. Henson P. M. Release of vasoactive amines from rabbit platelets induced by sensitized mononuclear leukocytes and antigen. J Exp Med. 1970 Feb;131(2):287–306. doi: 10.1084/jem.131.2.287. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kaliner M., Austen K. F. A sequence of biochemical events in the antigen-induced release of chemical mediators from sensitized human lung tissue. J Exp Med. 1973 Nov 1;138(5):1077–1094. doi: 10.1084/jem.138.5.1077. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Morgan P. H., Robinson N. C., Walsh K. A., Neurath H. Inactivation of bovine trypsinogen and chymotrypsinogen by diisopropylphosphorofluoridate. Proc Natl Acad Sci U S A. 1972 Nov;69(11):3312–3316. doi: 10.1073/pnas.69.11.3312. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Pearlman D. S., Ward P. A., Becker E. L. The requirement of serine esterase function in complement-dependent erythrophagocytosis. J Exp Med. 1969 Oct 1;130(4):745–764. doi: 10.1084/jem.130.4.745. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Ranadive N. S., Muir J. D. Similarity in the mechanism of histamine release induced by cationic protein from neutrophils and by complement dependent Ag-Ab reaction. Int Arch Allergy Appl Immunol. 1972;42(2):236–249. doi: 10.1159/000230609. [DOI] [PubMed] [Google Scholar]
  19. Trinchieri G., De Marchi M. Antibody-dependent cell-mediated cytotoxicity in humans. III. Effect of protease inhibitors and substrates. J Immunol. 1976 Apr;116(4):885–891. [PubMed] [Google Scholar]
  20. Ward P. A., Becker E. L. Mechanisms of the inhibition of chemotaxis by phosphonate esters. J Exp Med. 1967 Jun 1;125(6):1001–1020. doi: 10.1084/jem.125.6.1001. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Ward P. A., Becker E. L. The deactivation of rabbit neutrophils by chemotactic factor and the nature of the activatable esterase. J Exp Med. 1968 Apr 1;127(4):693–709. doi: 10.1084/jem.127.4.693. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from The Journal of Experimental Medicine are provided here courtesy of The Rockefeller University Press

RESOURCES