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Functional reconstitution of purified muscarinic receptors and inhibitory guanine nucleotide regulatory protein

Nature volume 316, pages 731–733 (1985)Cite this article

Abstract

Muscarinic receptors trigger several different responses including an increase in concentration of cyclic GMP, a decrease in cyclic AMP concentration, breakdown of polyphosphoinositides and changes in ion permeability. It is not yet clear whether these reactions occur sequentially or independently and which directly coupled to the muscarinic receptor. Several lines of evidence indicate that muscarinic receptors in many, if not all, cell types are coupled to the inhibitory guanine nucleotide regulatory protein (Ni or Gi) of adenylate cyclase. To provide direct evidence for this coupling, we have reconstituted muscarinic receptors purified from porcine brain with Ni purified from rat brain in a phospholipid vesicle. Here, we report that the GTPase activity of Ni is stimulated by carbachol. This action is blocked by the simultaneous addition of atropine and is not observed when the Ni protein is ADP-ribosylated. We conclude that one function of the muscarinic receptor is the activation of Ni.

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Authors and Affiliations

  1. Department of Biochemistry, Hamamatsu University School of Medicine, Hamamatsu, 431-31, Japan

    Kazuko Haga, Tatsuya Haga & Arata Ichiyama

  2. Department of Physiological Chemistry, Faculty of Pharmaceutical Sciences, Hokkaido University, Sapporo, 060, Japan

    Toshiaki Katada, Hitoshi Kurose & Michio Ui

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  1. Kazuko Haga
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  2. Tatsuya Haga
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  4. Toshiaki Katada
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Haga, K., Haga, T., Ichiyama, A. et al. Functional reconstitution of purified muscarinic receptors and inhibitory guanine nucleotide regulatory protein. Nature 316, 731–733 (1985). https://doi.org/10.1038/316731a0

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