Reversible or irreversible modification of [3H]PAF binding on rabbit platelet membranes differentiates various PAF receptor antagonists.
[3H]Platelet-activating factor (PAF) binding to rabbit platelet membranes was examined before and after 20 min preincubation at 25 degrees 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 BN 52021, 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.
Domingo
M T
MT
Institut Henri Beaufour, Les Ulis, France.
Piro
F
F
Broquet
C
C
Auclair
E
E
Chabrier
P E
PE
Braquet
P
P
D000818:Animals; D001381:Azepines / Q000494:pharmacology; D001792:Blood Platelets / Q000378:metabolism*; D002462:Cell Membrane / Q000378:metabolism; D004148:Dioxolanes / Q000494:pharmacology; D004224:Diterpenes; D005663:Furans / Q000494:pharmacology; D046934:Ginkgolides; D007783:Lactones / Q000494:pharmacology; D015394:Molecular Structure; D010972:Platelet Activating Factor / Q000031:analogs & derivatives / Q000378:metabolism* / Q000494:pharmacology; D010980:Platelet Membrane Glycoproteins; D011726:Pyridinium Compounds / Q000494:pharmacology; D011806:Quinolinium Compounds / Q000494:pharmacology; D011817:Rabbits; D011956:Receptors, Cell Surface / Q000037:antagonists & inhibitors*; D043562:Receptors, G-Protein-Coupled; D014230:Triazoles / Q000494:pharmacology; D014316:Tritium
D001381:Azepines; D004148:Dioxolanes; D004224:Diterpenes; D005663:Furans; D046934:Ginkgolides; D007783:Lactones; C029271:O-deacetyl platelet activating factor; D010972:Platelet Activating Factor; D010980:Platelet Membrane Glycoproteins; D011726:Pyridinium Compounds; D011806:Quinolinium Compounds; D011956:Receptors, Cell Surface; D043562:Receptors, G-Protein-Coupled; D014230:Triazoles; C039351:platelet activating factor receptor; D014316:Tritium; C052518:WEB 2086; C072668:BN 52111; C072669:BN 52115; C047504:2,5-bis(3,4,5-trimethoxyphenyl)tetrahydrofuran; C045856:ginkgolide B