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. 1984 May;52(1):169–174.

Isolation of a platelet membrane protein which binds the platelet-activating factor 1-0-hexadecyl-2-acetyl-SN-glycero-3-phosphorylcholine.

F H Valone
PMCID: PMC1454573  PMID: 6325330

Abstract

The phospholipid platelet-activating factor 1-0-hexadecyl-2-acetyl-SN-glycero-3-phosphorylcholine (AGEPC) initiates platelet function by interacting specifically with 1399 +/- 498 (mean +/- SD) high-affinity membrane receptors per platelet. In studies designated to characterize the high affinity binding site, AGEPC-human serum albumin-Sepharose was employed to isolate a 180,000 mol. wt. protein from human platelet plasma membranes. Platelet plasma membranes were isolated by adsorption of sonicated human platelets to a column of wheat germ agglutinin-Sepharose and elution with N-acetyl-glucosamine. The plasma membranes were solubilized in 5% sodium dodecyl sulphate and applied to a column of AGEPC-human serum albumin-Sepharose. After washing the column extensively, the specifically bound material was eluted with a five-fold molar excess of AGEPC. Sodium dodecyl sulphate polyacrylamide gel electrophoresis of the eluted material revealed a single protein with an apparent molecular weight of 180,000. This protein was not recovered from solubilized platelet membranes when chromatography was performed with a column of human serum albumin-Sepharose lacking AGEPC. The capacity of this protein to bind AGEPC suggests that it represents a constituent of the human platelet receptor for AGEPC.

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Selected References

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  1. 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]
  2. Berndt M. C., Phillips D. R. Purification and preliminary physicochemical characterization of human platelet membrane glycoprotein V. J Biol Chem. 1981 Jan 10;256(1):59–65. [PubMed] [Google Scholar]
  3. Cheng C. M., Hawiger J. Affinity isolation and characterization of immunoglobulin G Fc fragment-binding glycoprotein from human blood platelets. J Biol Chem. 1979 Apr 10;254(7):2165–2167. [PubMed] [Google Scholar]
  4. Demopoulos C. A., Pinckard R. N., Hanahan D. J. Platelet-activating factor. Evidence for 1-O-alkyl-2-acetyl-sn-glyceryl-3-phosphorylcholine as the active component (a new class of lipid chemical mediators). J Biol Chem. 1979 Oct 10;254(19):9355–9358. [PubMed] [Google Scholar]
  5. Goetzl E. J., Derian C. K., Tauber A. I., Valone F. H. Novel effects of 1-O-hexadecyl-2-acyl-sn-glycero-3-phosphorycholine mediators on human leukocyte function: delineation of the specific roles of the acyl substituents. Biochem Biophys Res Commun. 1980 Jun 16;94(3):881–888. doi: 10.1016/0006-291x(80)91317-0. [DOI] [PubMed] [Google Scholar]
  6. McManus L. M., Hanahan D. J., Pinckard R. N. Human platelet stimulation by acetyl glyceryl ether phosphorylcholine. J Clin Invest. 1981 Mar;67(3):903–906. doi: 10.1172/JCI110108. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Nachman R. L., Kinoshita T., Ferris B. Structural analysis of human platelet membrane glycoprotein I complex. Proc Natl Acad Sci U S A. 1979 Jun;76(6):2952–2954. doi: 10.1073/pnas.76.6.2952. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. O'Flaherty J. T., Lees C. J., Miller C. H., McCall C. E., Lewis J. C., Love S. H., Wykle R. L. Selective desensitization of neutrophils: further studies with 1-O-alkyl-sn-glycero-3-phosphocholine analogues. J Immunol. 1981 Aug;127(2):731–737. [PubMed] [Google Scholar]
  9. Rotman A., Linder S. Isolation of platelet membrane fractions by lectin affinity chromatography. Thromb Res. 1981 Apr 1;22(1-2):227–232. doi: 10.1016/0049-3848(81)90325-x. [DOI] [PubMed] [Google Scholar]
  10. Valone F. H., Coles E., Reinhold V. R., Goetzl E. J. Specific binding of phospholipid platelet-activating factor by human platelets. J Immunol. 1982 Oct;129(4):1637–1641. [PubMed] [Google Scholar]
  11. Valone F. H., Goetzl E. J. Enhancement of human polymorphonuclear leukocyte adherence by the phospholipid mediator 1-O-hexadecyl-2-acetyl-sn-glycero-3-phosphorylcholine (AGEPC). Am J Pathol. 1983 Oct;113(1):85–89. [PMC free article] [PubMed] [Google Scholar]
  12. Valone F. H., Goetzl E. J. Specific binding by human polymorphonuclear leucocytes of the immunological mediator 1-O-hexadecyl/octadecyl-2-acetyl-sn-glycero-3-phosphorylcholine. Immunology. 1983 Jan;48(1):141–149. [PMC free article] [PubMed] [Google Scholar]
  13. Valone F. H., Whitmer D. I., Pickett W. C., Austen K. F., Goetzl E. J. The immunological generation of a platelet-activating factor and a platet-lytic factor in the rat. Immunology. 1979 Aug;37(4):841–848. [PMC free article] [PubMed] [Google Scholar]
  14. Vargaftig B. B., Chignard M., Benveniste J., Lefort J., Wal F. Background and present status of research on platelet-activating factor (PAF-acether). Ann N Y Acad Sci. 1981;370:119–137. doi: 10.1111/j.1749-6632.1981.tb29727.x. [DOI] [PubMed] [Google Scholar]

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