Calcium channel subtypes in rat brain: biochemical characterization of the high-affinity receptors for omega-conopeptides SNX-230 (synthetic MVIIC), SNX-183 (SVIB), and SNX-111 (MVIIA).
High-threshold voltage-sensitive calcium channels of the N-type, L-type, and P-type have been distinguished in the mammalian CNS predominantly on the basis of their sensitivity to selective antagonists. Matching them with genes identified by molecular cloning is an ongoing undertaking. Whereas L-type channels are characterized by their sensitivity to dihydropyridines and P-type channels by sensitivity to the funnel-web spider toxin AgaIVA, the N-type channel has been shown to be recognized by the omega-conopeptides GVIA and MVIIA. Recently, two new members of the family of omega-conopeptides--MVIIC from the marine snail Conus magus and SVIB from Conus striatus--have been described. Binding and electrophysiological data suggest that these two peptides, in addition to interacting with N-type calcium channels, interact with a widely distributed receptor in neuronal membranes that is distinct from N-type channels. In this report we demonstrate through biochemical and pharmacological differentiation at individual receptor polypeptide resolution, by affinity cross-linking, SDS-PAGE, and autoradiography, that SNX-230 (synthetic MVIIC) binds with high affinity to a calcium channel alpha 1 subunit distinct from the high-affinity alpha 1 target of SNX-111 (synthetic MVIIA). SNX-183 (synthetic SVIB) interacts with both alpha 1 subunits with lower affinity. Whereas the alpha 1 subunit recognized with high affinity by MVIIA corresponds to the N-type channel, the other represents a novel calcium channel distinct from N-, L-, and perhaps P-type channels.
Woppmann
A
A
Neurex Corporation, Menlo Park, California 94025-1012.
Ramachandran
J
J
Miljanich
G P
GP
D000818:Animals; D001667:Binding, Competitive; D001923:Brain Chemistry; D015220:Calcium Channels / Q000145:classification* / Q000378:metabolism; D008297:Male; D008970:Molecular Weight; D009419:Nerve Tissue Proteins / Q000145:classification* / Q000378:metabolism; D010455:Peptides / Q000378:metabolism*; D011499:Protein Processing, Post-Translational; D051381:Rats; D017207:Rats, Sprague-Dawley; D013574:Synaptosomes / Q000378:metabolism; D020960:omega-Conotoxins
D015220:Calcium Channels; D009419:Nerve Tissue Proteins; D010455:Peptides; D020960:omega-Conotoxins; C077380:omega-conotoxin SVIB; C075851:omega-conotoxin-MVIIC; C078452:ziconotide