Towards the Crystal Structure Determination of Muscarinic Acetylcholine Receptors

  • Ryoji Suno
  • Hidetsugu Asada
  • Takuya Kobayashi
Part of the Neuromethods book series (NM, volume 107)


G protein-coupled receptors (GPCRs) constitute the largest family of receptors encoded by the human genome. Activation and inhibition of GPCRs under the physiological and pathophysiological conditions is largely mediated by chemical ligands (agonists and antagonists) that bind to the orthosteric binding pocket. Orthosteric ligands are, however, often nonspecific, binding to more than one GPCR subtype. In contrast to orthosteric agonists and antagonists, allosteric ligands do not directly compete with hormones and neurotransmitters for binding to the orthosteric binding pocket. Furthermore, allosteric ligands typically occupy structurally diverse regions of receptors and therefore are more selective for specific GPCRs, regulating receptor function in the more subtle ways by either enhancing or diminishing responses to natural ligands such as hormones or neurotransmitters. Recent X-ray crystallographic studies have provided detailed structural information regarding the nature of the orthosteric muscarinic binding site and an outer receptor cavity that can bind allosteric drugs. These new findings may guide the development of selective muscarinic receptor. The procedures involved in the production, purification, and crystallization of GPCRs are introduced here and facilitate a greater understanding of the structural basis of GPCR function.

Key words

G-protein coupled receptor X-ray crystal structure analysis Muscarinic acetylcholine receptor Antagonist Agonist Orthosteric binding site Allosteric binding site 



This work was supported by the Exploratory Research for Advanced Technology (ERATO) program of the Japan Science and Technology Agency (JST) (to T.K.), by the Toray Science Foundation (to T.K.), by Takeda Science Foundation (to T.K., R.S., and H.A.), by Ichiro Kanehara Foundation (to T.K.), by The Sumitomo Foundation (to T.K.), by the Core Research for Evolutional Science and Technology (CREST) program of the JST (to T.K.), and by the Platform for Drug Discovery, Informatics, and Structural Life Science from the Ministry of Education, Culture, Sports, Science and Technology, Japan (to T.K.).

Supplementary material

323171_1_En_1_MOESM1_ESM.pptx (5.6 mb)
Supplementary Fig. 1 This slide shows that structure of orthosteric domains similar among M1–M5 receptor subtypes. Asp103 conserved among all amine receptor has made a salt bridge with the amine of QNB, muscarinic receptor antagonist. Asn404 conserved among all muscarinic receptors has made hydrogen bonds with hydroxyl and carbonyl of QNB. There are 14 amino acids around QNB. Except Phe181, 13 amino acids of receptor are conserved among M1–M5 receptor subtypes. To develop the subtype specific ligand, we have to target the allosteric domain, not the orthosteric domain (PPTX 5750 kb)


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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • Ryoji Suno
    • 1
    • 3
  • Hidetsugu Asada
    • 1
  • Takuya Kobayashi
    • 1
    • 2
    • 3
  1. 1.Department of Medical Chemistry and Cell BiologyKyoto University Graduate School of MedicineKyotoJapan
  2. 2.Japan Science and Technology Agency (JST)Core Research for Evolutional Science and Technology (CREST)KyotoJapan
  3. 3.Platform for Drug Discovery, Informatics and Structural Life ScienceKyotoJapan

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