Characteristics of [3H]nimodipine binding to sarcolemmal membranes from rat vas deferens and its regulation by guanine nucleotide.
The binding properties of a 1,4-dihydropyridine (DHP) calcium entry blocker, [3H]nimodipine, to a microsomal fraction from rat vas deferens was characterized. The specific binding was saturable, rapid and reversible. Scatchard analysis of the binding revealed a single binding site, and the dissociation constant and the maximum number of binding sites were 0.31 +/- 0.02 nM and 97.0 +/- 7.19 fmol/mg protein, respectively. Both the Kd value obtained from the kinetic study and the IC50 value from relaxation of the K+-depolarized organ were approximately 0.4 nM, indicating that the binding site is closely related to the functional Ca2+ channel. The specific [3H]nimodipine binding was displaced by DHP derivatives at low concentration and by verapamil at high concentration, but diltiazem had no effect on the binding. Calcium chelating agents decreased the [3H]nimodipine binding which was restored by adding Ca2+. 5'-Guanylylimidodiphosphate caused a rightward shift of the displacement curve for Bay K 8644 but not for nimodipine, suggesting the involvement of guanine nucleotide binding protein in the signal transduction between the DHP binding site and the Ca2+ channel.
Higo
K
K
Department of Chemical Pharmacology, Faculty of Pharmaceutical Sciences, University of Tokyo, Japan.
Saito
H
H
Matsuki
N
N
D000818:Animals; D001665:Binding Sites; D015220:Calcium Channels / Q000378:metabolism; D002614:Chelating Agents / Q000494:pharmacology; D006165:Guanylyl Imidodiphosphate / Q000494:pharmacology; D066298:In Vitro Techniques; D007700:Kinetics; D008297:Male; D009126:Muscle Relaxation / Q000187:drug effects; D009553:Nimodipine / Q000378:metabolism* / Q000494:pharmacology; D051381:Rats; D011919:Rats, Inbred Strains; D012508:Sarcolemma / Q000378:metabolism; D014649:Vas Deferens / Q000187:drug effects / Q000378:metabolism*
D015220:Calcium Channels; D002614:Chelating Agents; D006165:Guanylyl Imidodiphosphate; D009553:Nimodipine