Crystal structure of an ACh-binding protein reveals the ligand-binding domain of nicotinic receptors

Brejc, Katjuša; Dijk, Willem J. van; Klaassen, Remco V.; Schuurmans, M.; Oost, John van der; Smit, August B.; Sixma, Titia K.

doi:10.1038/35077011

Cited by 1,705 publications

(2,066 citation statements)

References 47 publications

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“…Nevertheless, a5 di ers from typical a subunits in the domains thought to be important for agonist binding, lacking, for instance, Tyr93 and Tyr190. In nicotinic ACh receptors (nAChR) typical a subunits contribute either the whole of the agonist binding domain (Unwin, 2001) or the subsite for the ACh ester moiety (Karlin & Akabas, 1995;Brejc et al, 2001). A typical neuronal a subunit (a2, a3 or a4) will form functional nAChRs when expressed heterologously together with a typical b subunit (b2 or b4; Lindstrom, 2000).…”

Section: Introductionmentioning

confidence: 99%

Formation of functional α3β4α5 human neuronal nicotinic receptors in Xenopus oocytes: a reporter mutation approach

Groot-Kormelink

Boorman

Sivilotti

2001

British J Pharmacology

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1 The a5 subunit participates to the formation of native neuronal nicotinic receptors, particularly in autonomic ganglia. Like the related b3 subunit, a5 forms functional recombinant receptors if expressed together with a pair of typical a and b subunits, but its e ect on the properties of the resulting aba5 receptor depends on the a and b subunits chosen and on the expression system. We used a reporter mutation approach to test whether a5, like b3, is incorporated as a single copy in human a3b4a5 receptors expressed in oocytes. 2 As previously reported, the main indication of the presence of a5 in a3b4a5 wt was an increase in apparent receptor desensitization (compared with a3b4 receptors). If the a3b4a5 receptor bore a 9'T mutation in the second transmembrane domain of either a3 or b4, a5 incorporation produced a decrease in ACh sensitivity (by 4 fold for a3 LT b4a5 vs a3 LT b4 and by 40 fold for a3b4 LT a5 vs a3b4 LT ). The much greater e ect observed in a3b4 LT a5 receptors accords with the hypothesis that a5 takes the place of a b subunit in the receptor. 3 Introducing a 9'T mutation in a5 had no e ect on the agonist sensitivity of a3b4a5 receptors, but reduced apparent desensitisation, as judged by the sag in the current response to high agonist concentrations. 4 Introducing the 9'T mutation in a3 or b4 in the triplet receptor reduced the EC 50 for ACh by a similar extent (7 and 9 fold, respectively), suggesting that a3b4a5 receptors contain two copies each of a and b and therefore only one copy of a5.

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Section: Introductionmentioning

confidence: 99%

Formation of functional α3β4α5 human neuronal nicotinic receptors in Xenopus oocytes: a reporter mutation approach

Groot-Kormelink

Boorman

Sivilotti

2001

British J Pharmacology

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“…The initial crystal structure showed the amine containing buffer, HEPES at ~100mM concentration, to occupy the agonist site [6]. Crystallization in the presence of polyethylene glycol shows low occupation of the cryoprotectant in the agonist site and radial extension of the C loop from the core backbone of the molecule [11] (Fig.…”

Section: Crystal Structures Of Competitive Agonists and Antagonistsmentioning

confidence: 99%

“…Several structures of agonist (epibatidine, nicotine, lobeline, carbamylcholine) and antagonist (α-conotoxin, methyllycaconitine, α-bungarotoxin) bound to AChBP have been reported [6,[9][10][11][12], and other structures have been resolved at high resolution or are under study. Hence, one now has a fairly comprehensive perspective on structures of the AChBP complexes.…”

Section: Crystal Structures Of Competitive Agonists and Antagonistsmentioning

confidence: 99%

“…Sixma and colleagues have characterized pharmacologically and structurally a soluble protein, the acetylcholine binding protein (AChBP) from the fresh water snail, Lymnaea stagnalis, whose subunits are homologous to the extracellular domain of the nicotinic receptor. This soluble pentameric protein from Lymnaea, the related proteins from Bulinus, and the marine species, Aplysia, bind nicotinic ligands with the requisite specificity [6,7]. When the gene encoding an AChBP is coupled to a sequence encoding the transmembrane spans of the ligand-gated ion channel family, the expressed protein shows the capacity to gate ions when elicited by agonists, in a fashion expected for a receptor composed of the so derived expressed subunit chimera [8].…”

Section: Introductionmentioning

confidence: 99%

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Structure-guided drug design: Conferring selectivity among neuronal nicotinic receptor and acetylcholine-binding protein subtypes

Taylor

Talley

Radić

et al. 2007

Biochemical Pharmacology

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Neuronal nicotinic receptors, encoded by nine genes of the alpha and three of the beta type of subunits, and whose gene products assemble in distinct permutations as pentameric molecules, constitute a fertile area for structure-guided drug design. Design strategies are augmented by a wide variety of peptide, alkaloid and terpenoid toxins from various marine and terrestrial species that interact with nicotinic receptors. Also, acetylcholinebinding proteins from mollusks, as structural surrogates of the receptor that mimic its extracellular domain, provide atomic resolution templates for analysis of structure and response. Herein, we describe a structure-guided approach to nicotinic ligand design that employs crystallography of this protein as the basic template, but also takes into consideration the dynamic properties of the receptor molecules in their biological media. We present the crystallographic structures of several complexes of various agonists and antagonists that associate with the agonist site and can competitively block the action of acetylcholine. In so far as the extracellular domain is involved, we identify additional noncompetitive sites at those subunit interfaces where agonists do not preferentially bind. Ligand association at these interface sites may modulate receptor function. Ligand binding is also shown by solution-based spectroscopic and spectrometric methods to affect the dynamics of discrete domains of the receptor molecule. The surrogate receptor molecules can then be employed to design ligands selective for receptor subtype through the novel methods of freeze-frame, click chemistry that uses the very structure of the target molecule as a template for synthesis of the inhibitor.

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“…Previous studies have indicated that the nicotinic binding site is located at the centre of the extracellular domain of the receptor and lies at the interface between  and β subunits, which are known as the principal (P) and complementary (C) components, respectively [48][49][50]. Several residue stretches referred to as loops A, B and C in the  subunit, and D, E and F in the β subunit shape the binding site (in the following the symbols "" and "β" will be used before the residue to indicate the subunit it belongs to, and the numbering refers to the 4 and β2 subunits in rat).…”

Section: Binding Mode Of Mdmamentioning

confidence: 99%

Molecular basis of the selective binding of MDMA enantiomers to the alpha4beta2 nicotinic receptor subtype: Synthesis, pharmacological evaluation and mechanistic studies

Llabrés

García-Ratés

Cristóbal-Lecina

et al. 2014

European Journal of Medicinal Chemistry

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The α4β2 nicotinic acetylcholine receptor (nAChR) is a molecular target of 3,4-methylenedioxymethamphetamine (MDMA), a synthetic drug also known as ecstasy, and it modulates the MDMA-mediated reinforcing properties. However, the enantioselective preference of the α4β2 nAChR subtype still remains unknown. Since the two enantiomers exhibit different pharmacological profiles and stereoselective metabolism, the aim of this study is to assess a possible difference in the interaction of the MDMA enantiomers with this nAChR subtype. To this end, we report a novel simple, yet highly efficient enantioselective synthesis of the MDMA enantiomers, in which the key step is the diastereoselective reduction of imides derived from optically pure tert-butylsulfinamide. The enantioselective binding to the receptor is examined using

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Crystal structure of an ACh-binding protein reveals the ligand-binding domain of nicotinic receptors

Cited by 1,705 publications

References 47 publications

Formation of functional α3β4α5 human neuronal nicotinic receptors in Xenopus oocytes: a reporter mutation approach

Formation of functional α3β4α5 human neuronal nicotinic receptors in Xenopus oocytes: a reporter mutation approach

Structure-guided drug design: Conferring selectivity among neuronal nicotinic receptor and acetylcholine-binding protein subtypes

Molecular basis of the selective binding of MDMA enantiomers to the alpha4beta2 nicotinic receptor subtype: Synthesis, pharmacological evaluation and mechanistic studies

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