A photoaffinity ligand of the acetylcholine‐binding site predominantly labels the region 179–207 of the α‐subunit on native acetylcholine receptor from <i>Torpedo marmorata</i>

Dennis, Michael; Giraudat, Jérôme; Kotzyba‐Hibert, Florence; Goeldner, Maurice; Hirth, Christian; Chang, Jui‐Yoa; Changeux, Jean‐Pierre

doi:10.1016/0014-5793(86)81497-1

Cited by 47 publications

(19 citation statements)

References 27 publications

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“…Numerous lines of investigation on nicotinic acetylcholine receptors provide evidence that conserved clusters of aromatic amino acids are involved with ligand binding [92,[116][117][118][119][120][121][122][123][124][125]. Acetylcholinesterase also appears to have numerous aromatic amino acids in its binding gorge [126].…”

Section: Ii2 π-Cation and π-Aromatic Interactionmentioning

confidence: 99%

Exploring the Nature of Molecular Recognition in Nicotinic Acetylcholine Receptors

Schmitt

2000

CMC

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Nicotinic acetylcholine receptors (nAChRs) are the subject of ever increasing interest because of their presumed involvement in the etiology of numerous clinical disorders. Unfortunately, the absence of atomic-level structural data, as well as the pharmacological complexity of these receptors leaves many fundamental questions unanswered. An understanding of how ligands interact with the receptor and, in-turn, how these interactions lead to pharmacological effect is vital in the advancement of nAChR-based therapeutics. We will first explore physico-chemical themes that are evidenced to be of particular importance in nAChR molecular recognition; these are- pi-cation interaction, conformational entropy and stereochemistry. The second objective of this review is an interpretive encapsulation of the extensive and disparate body of structure-activity data that now exists for nAChRs. Finally, this review will advocate a re-investigation of distance geometry based methods as well as the need for additional approaches in nicotinic receptor pharmacophore generation.

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Section: Ii2 π-Cation and π-Aromatic Interactionmentioning

confidence: 99%

Exploring the Nature of Molecular Recognition in Nicotinic Acetylcholine Receptors

Schmitt

2000

CMC

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“…The use of p-N,N-dimethylammonium benzenediazonium difluoroborate (DDF), an affinity probe similar to TDF (3, 4), provided additional important information (56). The dimethylammonium group of DDF created a resonant molecule that could be photoactivated by energy transfer from the protein.…”

Section: Identification Of the Ach-binding Sitesmentioning

confidence: 99%

The Nicotinic Acetylcholine Receptor: The Founding Father of the Pentameric Ligand-gated Ion Channel Superfamily

Changeux

2012

Journal of Biological Chemistry

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211

149

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A critical event in the history of biological chemistry was the chemical identification of the first neurotransmitter receptor, the nicotinic acetylcholine receptor. Disciplines as diverse as electrophysiology, pharmacology, and biochemistry joined together in a unified and rational manner with the common goal of successfully identifying the molecular device that converts a chemical signal into an electrical one in the nervous system. The nicotinic receptor has become the founding father of a broad family of pentameric membrane receptors, paving the way for their identification, including that of the GABA A receptors.It has been 42 years since the isolation of the nicotinic acetylcholine receptor (nAChR) 2 from fish electric organ, the first ligand-gated ion channel and the first ion channel ever identified; 25 years since the first GABA A and glycine receptor subunits were cloned and sequenced and concomitantly their homology with the nAChRs recognized; and 5 years since the discovery that closely homologous ligand-gated ion channels are present in prokaryotes (1). In this minireview, I briefly retrace the main steps in the discovery of the nAChR, the titular head of this receptor superfamily. The Concept of Receptor and the Chemical Identification of the Acetylcholine ReceptorThe English physiologist John Newport Langley, working with neuromuscular preparations, proposed in 1905 that muscle tissue possesses "a substance that combines with nicotine and curare . . . receives the stimulus and transmits it." He called the muscle entity the "receptive substance." In the subsequent 50 years, the concept of pharmacological receptors inspired three main lines of research: first, the pharmacological approach aimed at characterizing the specificity of the receptor site by using novel chemical ligands (e.g. the distinction between nicotinic and muscarinic AChRs by Sir Henry Dale); second, the electrophysiological approach exemplified by Bernard Katz and John Eccles aimed at understanding the ionic responses to endogenous neurotransmitter signals; and third, the chemical tradition aimed at the chemical identification of the receptor molecule(s).In the late 1960s, lipids, polysaccharides, proteins, and even nucleic acids were considered as potential receptors. The early independent efforts of Carlos Chagas, Eduardo de Robertis, and David Nachmansohn to identify the receptor for acetylcholine (ACh) in the electric organ of the fish Electrophorus electricus with radioactive ligands were abandoned because their tissue extracts lacked specificity (2). However, in the course of these studies, Nachmansohn recognized the extraordinarily rich content of nicotinic synapses in the electric organ (2). With Ernest Schoffeniels, he devised a method for preparing individual cells, or electroplaques, from the electric organ. This offered the opportunity to investigate, simultaneously, the electrophysiological, pharmacological, and biochemical characteristics of the response to ACh within the same biological system (2). At this time, th...

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“…We report here the biologically active conformation of acetylcholine (AcCho) when bound to the high-affinity (desensitized) state ofthe acetylcholine receptor (AcChoR) from Torpedo californica. The exact location ofthe AcCho binding site on the a-subunits of the AcChoR is still unknown, although it has recently been localized to residues 158-216 by several groups using a variety oftechniques (1)(2)(3)(4). In contrast to the uncertainty in the AcCho binding site, crystal structures are available for AcCho and other agonists that also bind to the AcChoR (5-7).…”

mentioning

confidence: 99%

Conformation of acetylcholine bound to the nicotinic acetylcholine receptor.

Behling

Yamane²,

Navon³

et al. 1988

Proc. Natl. Acad. Sci. U.S.A.

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We report here the biologically active conformation ofacetylcholine when bound to the high-affinity state of the receptor from Torpedo cakfornica. The acetylcholine conformation was determined in the free and bound states by proton NMR two-dimensional nuclear Overhauser effects. In agreement with x-ray crystallographic data, acetylcholine in solution has an extended conformation with an average distance between the acetyl methyl and choline methyl protons of =5 A. When bound to the acetylcholine receptor, acetylcholine adopts a conformation where the acetyl methyl group is dose (3.3 A) to the methyl groups of the choline moiety. This bent conformation places the oxygens adjacent to one another and allows the methyl groups to form an uninterrupted hydrophobic surface over the rest of the acetylcholine molecule. The significant difference between the fre-and bound-state conformations implies that structure-activity studies based solely on molecular modeling strategies must be approached with caution. To understand the interactions of biologically interesting molecules, it is necessary to know the shape ofthe interaction sites as well as the conformations of the interacting molecules. We report here the biologically active conformation of acetylcholine (AcCho) when bound to the high-affinity (desensitized) state ofthe acetylcholine receptor (AcChoR) from Torpedo californica. The exact location ofthe AcCho binding site on the a-subunits of the AcChoR is still unknown, although it has recently been localized to residues 158-216 by several groups using a variety oftechniques (1-4). In contrast to the uncertainty in the AcCho binding site, crystal structures are available for AcCho and other agonists that also bind to the AcChoR (5-7). However, we show here that the conformation of AcCho in its receptor-bound state is distinctly different from its conformation in solution and in the crystalline solid state. MATERIALS AND METHODSAll experiments used purified AcChoR in asolectin vesicles. The receptor was purified from the electroplax organs of T. californica as described (8). We have shown that under the conditions used in this study, AcCho binds to the AcChoR and that selective proton relaxation measurements are very sensitive to this binding. [Under these conditions the ratio of Kd (acetylcholine)/Kd (nicotine) is 0.14.] Conditions (see text) have been determined to distinguish between the contributions from site-specific binding ofAcCho to the AcChoR and contributions from nonspecific interactions of AcCho both with the lipid and with other areas of the receptor (8).All proton NMR spectra were acquired on a 360-MHz Bruker AM series spectrometer with an Aspect 3000 computer. The two-dimensional nuclear Overhauser effect (NOE) data sets were obtained in the phase-sensitive mode (9) and were processed with Dennis Hare's Fourier transform NMR program. The temperature in all experiments was held at 250C to within PC with the Bruker variable temperature unit by passing heated nitrogen gas through the probe.The conf...

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A photoaffinity ligand of the acetylcholine‐binding site predominantly labels the region 179–207 of the α‐subunit on native acetylcholine receptor from Torpedo marmorata

Cited by 47 publications

References 27 publications

Exploring the Nature of Molecular Recognition in Nicotinic Acetylcholine Receptors

Exploring the Nature of Molecular Recognition in Nicotinic Acetylcholine Receptors

The Nicotinic Acetylcholine Receptor: The Founding Father of the Pentameric Ligand-gated Ion Channel Superfamily

Conformation of acetylcholine bound to the nicotinic acetylcholine receptor.

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