Inhibition of the type 1 inositol 1,4,5-trisphosphate-sensitive Ca2+ channel by calmodulin antagonists.
This study describes the effects of a number of calmodulin antagonists on the cerebellar type 1 inositol 1,4,5-trisphosphate (InsP3) receptor. All the antagonists tested (trifluoperazine, fluphenazine, chlorpromazine and calmidazolium) inhibited the extent of InsP3-induced Ca2+ release (IICR) with similar IC(50) values (between 60 and 85 microM). They did not affect the efficacy of InsP3 to release Ca2+, since the concentrations of InsP3 required to cause half-maximal release was little affected in the presence of these agents. In addition, these agents did not affect InsP3 binding to its receptor. Stopped-flow studies to determine the rate constants of IICR showed this process to be biphasic with a fast and slow component. All the calmodulin antagonists appeared to reduce the rate constants for Ca2+ release in a phase-specific manner, preferentially reducing the fast phase component. Chlorpromazine (75 microM) appeared to have the most potent effect on the fast phase rate constant, reducing it from 1.0 to 0.08 s(-1), while only reducing the rate constant for the slow phase about twofold (0.2-0.08 s(-1)). The fact that calmodulin itself inhibits both IICR and InsP3 binding, while these calmodulin antagonists also reduce Ca2+ release and do not affect InsP3 binding, suggests that the mechanism of action of these agents is unlikely to be due to the reversal of the modulatory action of calmodulin on this receptor.
School of Biosciences, University of Birmingham, Edgbaston, B15 2TT, Birmingham, UK.
S Z
SZ
Khan
J L
JL
Dyer
F
F
Michelangeli
D000818:Animals; D015220:Calcium Channels / Q000187:drug effects*; D002147:Calmodulin / Q000037:antagonists & inhibitors*; D002531:Cerebellum / Q000166:cytology*; D002746:Chlorpromazine; D004305:Dose-Response Relationship, Drug; D005476:Fluphenazine; D007093:Imidazoles; D066298:In Vitro Techniques; D053496:Inositol 1,4,5-Trisphosphate Receptors; D008861:Microsomes / Q000187:drug effects / Q000378:metabolism; D010640:Phenothiazines / Q000008:administration & dosage*; D018160:Receptors, Cytoplasmic and Nuclear / Q000037:antagonists & inhibitors*; D013552:Swine; D014268:Trifluoperazine
D015220:Calcium Channels; D002147:Calmodulin; D007093:Imidazoles; D053496:Inositol 1,4,5-Trisphosphate Receptors; D010640:Phenothiazines; D018160:Receptors, Cytoplasmic and Nuclear; D014268:Trifluoperazine; C031938:calmidazolium; D005476:Fluphenazine; D002746:Chlorpromazine