A monoclonal antibody to the beta subunit of the skeletal muscle dihydropyridine receptor immunoprecipitates the brain omega-conotoxin GVIA receptor

Junshi Sakamoto; Kevin P Campbell

doi:10.1016/S0021-9258(18)55150-4

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A monoclonal antibody to the beta subunit of the skeletal muscle dihydropyridine receptor immunoprecipitates the brain omega-conotoxin GVIA receptor

Journal article

Open access

Peer reviewed

A monoclonal antibody to the beta subunit of the skeletal muscle dihydropyridine receptor immunoprecipitates the brain omega-conotoxin GVIA receptor

Junshi Sakamoto and Kevin P Campbell

The Journal of biological chemistry, Vol.266(28), pp.18914-18919

10/05/1991

DOI: 10.1016/S0021-9258(18)55150-4

PMID: 1655767

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Abstract

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.

Rabbits

Calcium Channels

Immunoblotting

Peptides, Cyclic - immunology

Receptors, Neurotransmitter - analysis

Calcium Channel Blockers - pharmacology

Peptides, Cyclic - metabolism

Precipitin Tests

Receptors, Nicotinic - immunology

Animals

Receptors, Nicotinic - analysis

Sensitivity and Specificity

Muscles - chemistry

Receptors, Neurotransmitter - immunology

Brain Chemistry

Antibodies, Monoclonal - immunology

Conotoxins

Details

Title: Subtitle: A monoclonal antibody to the beta subunit of the skeletal muscle dihydropyridine receptor immunoprecipitates the brain omega-conotoxin GVIA receptor
Creators: Junshi Sakamoto - Howard Hughes Medical Institute, Department of Physiology and Biophysics, University of Iowa College of Medicine, Iowa City 52242
Kevin P Campbell
Resource Type: Journal article
Publication Details: The Journal of biological chemistry, Vol.266(28), pp.18914-18919
DOI: 10.1016/S0021-9258(18)55150-4
PMID: 1655767
NLM abbreviation: J Biol Chem
ISSN: 0021-9258
eISSN: 1083-351X
Publisher: United States
Grant note: HL39265 / NHLBI NIH HHS HL14388 / NHLBI NIH HHS
Language: English
Date published: 10/05/1991
Academic Unit: Neurology; Molecular Physiology and Biophysics; Iowa Neuroscience Institute
Record Identifier: 9984068370602771

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