Affinity chromatography of the muscarinic acetylcholine receptor.

作者: K Haga , T Haga

DOI: 10.1016/S0021-9258(17)43953-6

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摘要: Abstract A novel compound, 3-(2'-aminobenzhydryloxy)-tropane (ABT), and an ABT-agarose gel were synthesized used for the purification of solubilized muscarinic receptors. ABT had a high affinity with apparent dissociation constant (Kd) 7 nM receptors from porcine brain by digitonin. An was prepared coupling epoxy-activated Sepharose 6B, degree substitution to determined be 4-5 mumol/ml UV absorption spectrum. During chromatography using 10 ml 100 digitonin-solubilized preparation, 70% adsorbed gel, in marked contrast adsorption only 2% proteins. Approximately 25% applied eluted biospecifically 1 mM ligands. The purified fraction showed [3H]quinuclidinyl benzylate Kd 0.4 similar specificity ligands that unpurified soluble protein concentration too low accurately, but very approximately 10(3)-fold indicated.

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