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Reversible or irreversible modification of [3H]PAF binding on rabbit platelet membranes differentiates various PAF receptor antagonists

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Lipids

Abstract

[3H]Platelet-activating factor (PAF) binding to rabbit platelet membranes was examined before and after 20 min preincubation at 25°C in the presence of PAF, lysoPAF, or of five different PAF receptor antagonists (L 652731, BN 52021, WEB 2086, BN 52111 and BN 52115). When platelet membranes were not washed after preincubation with PAF or PAF antagonists, no significant specific binding of [3H]PAF was observed, which suggests full occupancy of the binding sites. When membranes were extensively washed, full recovery of specific [3H]PAF binding was attained with L 652731 and partial recoveries (60%, 55% and 30%) were reached with BN52021, WEB 2086 and PAF, respectively; no recovery was seen with the dioxolanes BN 52111 and BN 52115. Scatchard analysis of the binding data indicated that no significant change in the dissociation constant (Kd) and maximum number of binding sites (Bmax) occurred after preincubation of platelet membrane with L 652731, whereas a reduction of Bmax was observed when PAF and BN 52021 were measured. When platelet membranes were preincubated with WEB 2086, Bmax and Kd significantly increased. The data suggest differing binding properties for PAF and the PAF antagonists. Some of the PAF antagonists may tightly bind to the PAF receptor site(s) and/or irreversibly modify or downregulate PAF recognition sites. Our results also suggest that the interaction of PAF receptor antagonists with PAF receptor can be divided into at least two components, namely a reversible component and an irreversible one.

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Abbreviations

BSA:

bovine serum albumin

DMSO:

dimethylsulfoxide

EDTA:

ethylenediaminetetraacetic acid

lysoPAF:

1-O-alkyl-sn-glycero-3-phosphocholine

PAF:

platelet-activating factor, 1-O-alkyl-2-acetyl-sn-glycero-3-phosphocholine

PMSF:

phenylmethylsulfonyl fluoride

PRP:

platelet-rich plasma

References

  1. Demopoulos, C.A., Pinckard, R.N., and Hanahan, D.J. (1979)J. Biol. Chem. 254, 9355–9358.

    PubMed  CAS  Google Scholar 

  2. Benveniste, J., Tence, M., Varence, P., Bidault, J., Boulet, C., and Polonsky, J. (1979)C.R. Hebd. Seances. Acad. Sci. Ser. D. Sc. Nat. 289, 1037–1040.

    CAS  Google Scholar 

  3. Vargaftig, B.B., and Benveniste, J. (1983)Trends Pharmacol. Sci. 4, 341–344.

    Article  CAS  Google Scholar 

  4. Pinckard, R.N., McManus, L.N., Halonen, M., and Hanahan, D.J. (1983) inImmuno-pharmacology of the Lung (Newball, H.H., ed.) pp. 73–107, Marcel Decker, New York.

    Google Scholar 

  5. Benveniste, J., Boulet, C., Brink, C., and Labat, C. (1983)Br. J. Pharmacol. 80, 81–83.

    PubMed  CAS  Google Scholar 

  6. Baranes, J., Hellegouarch, A., Le Hegarat, M., Viossat, I., Auguet, M., Chabrier, P.E., Clostre, F., and Braquet, P. (1986)Pharmacol. Res. Commun. 18, 717–737.

    Article  PubMed  Google Scholar 

  7. Pirotzky, E., and Bidault, J. (1983) inPlatelet-Activating Factor and Structurally Related Ether-Lipids, Vol. 23 (Benveniste, J., and Arnoux, B., eds.) Elsevier Science Publishers, Amsterdam, 117–122.

    Google Scholar 

  8. Braquet, P., Spinnewyn, B., Braquet, M., Bourgain, R.H., Taylor, J.E., Etienne, A., and Drieu, K. (1985)Blood Vessels 16, 659–572.

    Google Scholar 

  9. Hwang, S.B., Lee, S.C., Cheah, J., and Shen, T.Y. (1983)Biochemistry 22, 4756–4763.

    Article  PubMed  CAS  Google Scholar 

  10. Inarrea, P., Gomez-Cambronero, J., Nieto, N., and Sanchez-Crespo, M. (1984)Eur. J. Pharmacol. 105, 309–315.

    Article  PubMed  CAS  Google Scholar 

  11. Domingo, M.T., Chabrier, P.E., Thiberghien, C., Wisner, A., Dray, F., and Braquet, P. (1988) inGinkgolides: Chemistry, Biology, Pharmacology and Clinical Perspectives, (Braquet, P. ed.) pp. 79–84, J.R. Prous Science Publishers, Barcelona.

    Google Scholar 

  12. Hwang, S.B., Lam, M.H., and Shen, T.Y. (1985)Biochem. Biophys. Res. Commun. 128, 972–979.

    Article  PubMed  CAS  Google Scholar 

  13. Bussolino, F., Tetta, C., and Camussi, G. (1984)Agents and Actions 15, 1–2.

    Article  Google Scholar 

  14. O'Flaherty, J.T., Surles, J.R., Redman, J., Jacobson, D., Piantadosi, C., and Wykle, R.L. (1986)J. Clin. Invest. 78, 381–388.

    Article  PubMed  Google Scholar 

  15. Hwang, S.B., (1987)Arch. Biochem. Biophys. 257, 339–344.

    Article  PubMed  CAS  Google Scholar 

  16. Domingo, M.T., Spinnewyn, B., Chabrier, P.E., and Braquet, P. (1988)Biochem. Biophys. Res. Commun. 151, 730–736.

    Article  PubMed  CAS  Google Scholar 

  17. Junier, P.P., Thiberghien, C., Rougeot, C., Fafeur, V., and Dray, F. (1988)Endocrinology 123, 172–180.

    Article  Google Scholar 

  18. Domingo, M.T., Chabrier, P.E., Van Delft, J.L., Verbeij, N.L., Van Haeringen, N.J., and Braquet, P. (1989)Biochem. Biophys. Res. Commun. 160, 250–256.

    Article  PubMed  CAS  Google Scholar 

  19. Terashita, Z.I., Imura, Y., and Nishikawa, K. (1985)Biochem. Pharmacol. 34, 1491–1495.

    Article  PubMed  CAS  Google Scholar 

  20. Hwang, S.B., Lam, M.H., Biftu, T., Beatie, T.S., and Shen, T.Y. (1985)J. Biol. Chem. 260, 15639–15645.

    PubMed  CAS  Google Scholar 

  21. Casals-Stenzel, J., Muacevic, G., and Weber, K.H. (1987)J. Pharmacol. Exp. Ther. 241, 974–981.

    PubMed  CAS  Google Scholar 

  22. Broquet, C., Auclair, E., Blavet, N., Touvay, C. and Braquet, P. (1990)Eur. J. Med. Chem. 25, 235–240.

    Article  CAS  Google Scholar 

  23. Shen, T.Y., Hwang, S.B., Chang, M.N., Doebber, T.W., Lam, M.H.T. Wu, M.S., Wang, X., Han, G.Q., and Li, R.Z. (1985)Proc. Natl. Aca. Sci. USA 82, 672–676.

    Article  CAS  Google Scholar 

  24. Korth, R., Nunez, D., Braquet, P., and Benveniste, J. (1987)Eur. J. Pharmacol. 142, 331–341.

    Article  PubMed  CAS  Google Scholar 

  25. Kloprogge, E.D., and Akkerman, W. (1984)Biochem. J. 224, 901–903.

    Google Scholar 

  26. Valone, F.H., Coles, E., Reinhold, V.R., and Goetzl, E.J. (1982)J. Immunoo. 129, 1637–1641.

    CAS  Google Scholar 

  27. Hwang, S.B., Lam, M.H., and Pong, S.S. (1986)J. Biol. Chem. 261, 532–537.

    PubMed  CAS  Google Scholar 

  28. Bradford, M.H. (1976)Anal. Biochem. 72, 248.

    Article  PubMed  CAS  Google Scholar 

  29. Portoghese, P.S., Larson, D.L., Jiang, J.B., Takemori, A.E., and Caruso, T.P. (1978)J. Med. Chem. 21, 598–599.

    Article  PubMed  CAS  Google Scholar 

  30. Broquet, C., Etienne, A., Touvay, C., Pignol, B., Mencia-Huerta, J.M., and Braquet, P. (1991)C.R.C. Handbook of PAF and PAF Antagonists (Braquet, P., ed.) CRC Press, Boca Raton.

    Google Scholar 

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Authors and Affiliations

  1. Institut Henri Beaufour 1, A venue des Tropiques, 91952, Les Ulis Cedex, France

    M. T. Domingo, F. Piro, C. Broquet, E. Auclair, P. E. Chabrier & P. Braquet

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  1. M. T. Domingo
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  2. F. Piro
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  3. C. Broquet
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  4. E. Auclair
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  5. P. E. Chabrier
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  6. P. Braquet
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Additional information

Based on a paper presented at the Third International Conference on Platelet-Activating Factor and Structurally Related Alkyl Ether Lipids, Tokyo, Japan, May 1989.

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Domingo, M.T., Piro, F., Broquet, C. et al. Reversible or irreversible modification of [3H]PAF binding on rabbit platelet membranes differentiates various PAF receptor antagonists. Lipids 27, 582–586 (1992). https://doi.org/10.1007/BF02536114

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  • DOI: https://doi.org/10.1007/BF02536114

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