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Effects of trypsin-treatment on agonist binding affinity to the muscarinic acetylcholine receptor
Authors:Kazuo Matsumoto  Shuji Uchida  Kunio Takeyasu  Hiroshi Higuchi  Hiroshi Yoshida
Institution:Department of Pharmacology I, Osaka University School of Medicine Nakanoshima, Kita-ku, Osaka 530, Japan
Abstract:Trypsin-treatment of the microsome fraction of the ileum and the synaptic membrane fraction of the cerebral cortex of guinea-pig caused selective reduction in the apparent affinity of an agonist (carbachol), but not an antagonist (atropine), to muscarinic acetylcholine receptors (mAChR), measured as inhibition of binding of 3H-quinuclidinyl benzilate (3H-QNB). This effect was similar to that of Gpp(NH)p. The effects of trypsin and Gpp(NH)p were not additive. On the other hand, treatment of these fractions with 5,5'-dithiobis (2-nitrobenzoic acid) (DTNB) increased the apparent affinity of agonist, but not antagonist. The effect of DTNB predominated over those of trypsin and Gpp(NH)p, when the fractions were treated with two reagents simultaneously.
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