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Isolation of the ω-conotoxin receptor from rabbit brain using affinity chromatography with a monoclonal antibody
Abstract   Open access

Isolation of the ω-conotoxin receptor from rabbit brain using affinity chromatography with a monoclonal antibody

Junshi Sakamoto and Kevin P. Campbell
Japanese journal of pharmacology, Vol.58(suppl.1), pp.230-230
1992
DOI: 10.1016/S0021-5198(19)49190-7
url
https://doi.org/10.1016/S0021-5198(19)49190-7View
Published (Version of record) Open Access

Abstract

There are several types of voltage-gated Ca channels. The N-type channels are blocked by ω-conotoxin GVIA (ωCgTX) and may be responsible for triggering neurotransmitter release. Although the skeletal muscle Ca channel has been already purified, the neuronal Ca channels have not, and their subunit compositions are still unknown. Since we have found that monoclonal antibody VD2_1 against the β subunit of the skeletal muscle Ca channel specifically immunoprecipitates the ωCgTX receptor. VD2_1 was utilyzed for isolation of the receptor. Rabbit brain membranes were preincubated with 「^^125 I」ωCgTX, solubilized with 1 % digitonin and vas first applied to a heparin-agarose column. The eluate from the column was then applied on a column of VD2_1 immobilized to agarose. The eluate with pH 10-buffer contained several proteins, some of which comigrated with the 「^^125 I」ωCgTX on a sucrose density gradient. Results of crosslinking, autoradiographic, and Western blotting experiments suggest a unique subunit composition of the brain ωCgTX receptor.

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