A monoclonal antibody to the beta subunit of the skeletal muscle dihydropyridine receptor immunoprecipitates the brain omega-conotoxin GVIA receptor.
Antibodies against the subunits of the dihydropyridine-sensitive L-type calcium channel of skeletal muscle were tested for their ability to immunoprecipitate the high affinity (Kd = 0.13 nM) 125I-omega-conotoxin GVIA receptor from rabbit brain membranes. Monoclonal antibody VD2(1) against the beta subunit of the dihydropyridine receptor from skeletal muscle specifically immunoprecipitated up to 86% of the 125I-omega-conotoxin receptor solubilized from brain membranes whereas specific antibodies against the alpha 1, alpha 2, and gamma subunits did not precipitate the brain receptor. Purified skeletal muscle dihydropyridine receptor inhibited the immunoprecipitation of the brain omega-conotoxin receptor by monoclonal antibody VD2(1). The dihydropyridine receptor from rabbit brain membranes was also precipitated by monoclonal antibody VD2(1). However, neither the neuronal ryanodine receptor nor the sodium channel was precipitated by monoclonal antibody VD2(1). The omega-conotoxin receptor immunoprecipitated by monoclonal antibody VD2(1) showed high affinity 125I-omega-conotoxin binding, which was inhibited by unlabeled omega-contoxin and by CaCl2 but not by nitrendipine or by diltiazem. An antibody against the beta subunit of the skeletal muscle dihydropyridine receptor stained 58- and 78-kDa proteins on immunoblot of the omega-conotoxin receptor, partially purified through heparin-agarose chromatography and VD2(1)-Sepharose chromatography. These results suggest that the brain omega-conotoxin-sensitive calcium channel contains a component homologous to the beta subunit of the dihydropyridine-sensitive calcium channel of skeletal muscle and brain.
Sakamoto
J
J
Howard Hughes Medical Institute, Department of Physiology and Biophysics, University of Iowa College of Medicine, Iowa City 52242.
Campbell
K P
KP
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D001923:Brain Chemistry; D002121:Calcium Channel Blockers / Q000494:pharmacology; D015220:Calcium Channels; D020916:Conotoxins; D015151:Immunoblotting; D009132:Muscles / Q000737:chemistry*; D010456:Peptides, Cyclic / Q000276:immunology / Q000378:metabolism; D011233:Precipitin Tests; D011817:Rabbits; D017981:Receptors, Neurotransmitter / Q000032:analysis* / Q000276:immunology; D011978:Receptors, Nicotinic / Q000032:analysis* / Q000276:immunology; D012680:Sensitivity and Specificity
D000911:Antibodies, Monoclonal; D002121:Calcium Channel Blockers; D015220:Calcium Channels; D020916:Conotoxins; D010456:Peptides, Cyclic; D017981:Receptors, Neurotransmitter; D011978:Receptors, Nicotinic; C053157:omega-conotoxin receptor; C055407:conotoxin GV