PURIFIED OMEGA-CONOTOXIN GVIA RECEPTOR OF RAT-BRAIN RESEMBLES A DIHYDROPYRIDINE-SENSITIVE L-TYPE CALCIUM-CHANNEL

被引:113
作者
MCENERY, MW
SNOWMAN, AM
SHARP, AH
ADAMS, ME
SNYDER, SH [1 ]
机构
[1] JOHNS HOPKINS MED INST, DEPT NEUROSCI, 725 N WOLFE ST, BALTIMORE, MD 21205 USA
[2] JOHNS HOPKINS MED INST, DEPT PHARMACOL & MOLEC SCI, BALTIMORE, MD 21205 USA
[3] JOHNS HOPKINS MED INST, DEPT PSYCHIAT & BEHAV SCI, BALTIMORE, MD 21205 USA
[4] UNIV CALIF RIVERSIDE, DEPT ENTOMOL, RIVERSIDE, CA 92521 USA
关键词
VOLTAGE-DEPENDENT CALCIUM CHANNELS; OMEGA-AGATOXIN IIIA; PHOTOAFFINITY LABELING;
D O I
10.1073/pnas.88.24.11095
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
The omega-conotoxin GVIA (CTX) receptor has been purified 1900-fold to apparent homogeneity by monitoring both reversible binding of I-125-labeled CTX (I-125-CTX) and photoincorporation of N-hydroxysuccinimidyl-4-azidobenzoate-I-125-CTX (HSA-I-125-CTX). Photoincorporation of HSA-I-125-CTX into a 230-kDa protein exhibits a pharmacologic and chromatographic profile indicating that the 230-kDa protein is the CTX-binding subunit of the receptor. The pharmacologic specificity of I-125-CTX binding to the purified CTX receptor closely resembles that of the native membrane-bound form with respect to sensitivity towards CTX (K(d) = 32 pM) and other peptide toxin antagonists. The purified CTX receptor comprises the 230-kDa protein (alpha-1) and four additional proteins with apparent molecular masses of 140 (alpha-2), 110, 70 (beta-2), and 60 (beta-1) kDa. This subunit structure closely resembles that of the 1,4-dihydropyridine-sensitive L-type calcium channel.
引用
收藏
页码:11095 / 11099
页数:5
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