Omega-conotoxin CVIB differentially inhibits native and recombinant N- and P/Q-type calcium channels.
Omega-conotoxins are routinely used as selective inhibitors of different classes of voltage-gated calcium channels (VGCCs) in excitable cells. In the present study, we examined the potent N-type VGCC antagonist omega-conotoxin CVID and non-selective N- and P/Q-type antagonist CVIB for their ability to block native VGCCs in rat dorsal root ganglion (DRG) neurons and recombinant VGCCs expressed in Xenopus oocytes. Omega-conotoxins CVID and CVIB inhibited depolarization-activated whole-cell VGCC currents in DRG neurons with pIC50 values of 8.12 +/- 0.05 and 7.64 +/- 0.08, respectively. Inhibition of Ba2+ currents in DRG neurons by CVID (approximately 66% of total) appeared to be irreversible for > 30 min washout, whereas Ba2+ currents exhibited rapid recovery from block by CVIB (> or = 80% within 3 min). The recoverable component of the Ba2+ current inhibited by CVIB was mediated by the N-type VGCC, whereas the irreversibly blocked current (approximately 22% of total) was attributable to P/Q-type VGCCs. Omega-conotoxin CVIB reversibly inhibited Ba2+ currents mediated by N- (Ca(V)2.2) and P/Q- (Ca(V)2.1), but not R- (Ca(V)2.3) type VGCCs expressed in Xenopus oocytes. The alpha2delta1 auxiliary subunit co-expressed with Ca(V)2.2 and Ca(V)2.1 reduced the sensitivity of VGCCs to CVIB but had no effect on reversibility of block. Determination of the NMR structure of CVIB identified structural differences to CVID that may underlie differences in selectivity of these closely related conotoxins. Omega-conotoxins CVIB and CVID may be useful as antagonists of N- and P/Q-type VGCCs, particularly in sensory neurons involved in processing primary nociceptive information.
Motin
Leonid
L
School of Biomedical Sciences, The University of Queensland, Brisbane, Queensland 4072, Australia.
Yasuda
Takahiro
T
Schroeder
Christina I
CI
Lewis
Richard J
RJ
Adams
David J
DJ
D000818:Animals; D000831:Animals, Newborn; D002121:Calcium Channel Blockers / Q000494:pharmacology*; D020864:Calcium Channels, N-Type / Q000502:physiology*; D020907:Calcium Channels, Q-Type / Q000502:physiology*; D004305:Dose-Response Relationship, Drug; D004347:Drug Interactions; D004558:Electric Stimulation; D005727:Ganglia, Spinal / Q000166:cytology; D008564:Membrane Potentials / Q000187:drug effects*; D008845:Microinjections / Q000379:methods; D008958:Models, Molecular; D009474:Neurons / Q000187:drug effects; D009865:Oocytes; D018408:Patch-Clamp Techniques; D011487:Protein Conformation; D051381:Rats; D014982:Xenopus laevis; D020960:omega-Conotoxins / Q000737:chemistry / Q000494:pharmacology*
D002121:Calcium Channel Blockers; D020864:Calcium Channels, N-Type; D020907:Calcium Channels, Q-Type; D020960:omega-Conotoxins