Molecular Genetics of Torpedo Marmorata Acetylcholine Receptor

Devillers‐Thiéry, Anne; Giraudat, Jérôme; Bentaboulet, Martine; Klarsfeld, André; Changeux, Jean Pierre

doi:10.1007/978-1-4684-4868-9_3

Cited by 7 publications

(6 citation statements)

References 34 publications

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“…californica (Noda et al, 1982(Noda et al, , 1983aClaudio et al, 1983), the a-chains of T. marmorata (Devillers-Thiery et al, 1983) and human and bovine muscle AChR (Noda et al, 1983b) revealed marked homologies among the different species. It has been proposed (Noda et al, 1982) that an antigenic site resides in the region of residues 161-166 of the a-chain of the Torpedo AChR.…”

mentioning

confidence: 96%

Synthesis of an antigenic site of native acetylcholine receptor peptide 159-169 of Torpedo acetylcholine receptor α-chain

McCormick¹,

Lennon²,

Atassi

1985

Biochemical Journal

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A region of the alpha-subunit of the nicotinic acetylcholine receptor (AChR) of the Torpedo electric organ, containing residues 161-166, has been proposed to be a major antigenic site in the native AChR protein. We report the synthesis of a peptide corresponding to residues 159-169, which contains the proposed antigenic region. In quantitative radiometric titrations, radiolabelled anti-(native AChR) antibodies from three different species, rabbit, rat and dog, exhibited considerable binding (approx. 15% relative to native AChR) to Sepharose-immobilized peptide 159-169, but did not bind significantly to Sepharose-immobilized unrelated proteins or peptides. Specificity was further confirmed by the finding that no rabbit anti-AChR antibodies bound to the peptide after absorption with native AChR. These data indicate that the region 159-169 contains an antigenic site that is readily accessible in solubilized native Torpedo AChR.

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confidence: 96%

Synthesis of an antigenic site of native acetylcholine receptor peptide 159-169 of Torpedo acetylcholine receptor α-chain

McCormick¹,

Lennon²,

Atassi

1985

Biochemical Journal

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“…californica have been sequenced (Noda et al, 1982(Noda et al, , 1983aClaudio et al, 1983). Based upon sequence analysis and structural topology of the a-subunit, it has been proposed that the invariant cysteine residues 128 and 142 comprise the disulphide bridge, the integrity of which is essential for the binding of acetylcholine to the receptor (Karlin, 1980;Noda et al, 1982Noda et al, , 1983bNoda et al, , 1983cDevillers-Thiery et al, 1983). We have undertaken studies to localize by peptide synthesis the acetylcholine binding site in both human and Torpedo receptors.…”

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confidence: 99%

Localization and synthesis of the acetylcholine-binding site in the α-chain of the Torpedo californica acetylcholine receptor

McCormick¹,

Atassi²

1984

Biochemical Journal

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The sequence of the alpha-chain of the acetylcholine receptor of T. californica has been determined by recent cloning studies. The integrity of the disulphide bond between Cys-128 and cys-142 has been shown to be important for the maintenance of the binding activity of the receptor, thus implicating the regions around the disulphide bridge in binding with acetylcholine. In the present work, a synthetic peptide containing this loop region (residues 125-147) was synthesized. Solid-phase radiometric binding assays demonstrated a high binding of 125I-labelled alpha-bungarotoxin to the synthetic peptide. It was further shown that the free peptide bound well to [3H]acetylcholine. Additional experiments demonstrated that pretreatment of peptide 125-147 with 2-mercaptoethanol destroyed its binding activity, clearly showing that the integrity of the disulphide structure was essential for binding. Unlabelled acetylcholine also inhibited the binding of labelled acetylcholine to the synthetic peptide. The region 125-147, therefore, contains essential elements of the acetylcholine binding site of the Torpedo receptor.

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“…The sequence of the ECD-Asu-ACR-16 and the human α7 nAChR chimera (SwissProt ID: P36544; Peng et al., 1994 ) were aligned using CLUSTALW multiple alignment ( Thompson et al., 1994 ). The sequence of the TID-Asu-ACR-16 and TID-Tma-nAChR (SwissProt ID: P02711; Devillers-Thiery et al., 1983 , Devillers-Thiery et al., 1984 ) were aligned using CLUSTALW.…”

Section: Methodsmentioning

confidence: 99%

The Ascaris suum nicotinic receptor, ACR-16, as a drug target: Four novel negative allosteric modulators from virtual screening

Zheng

Robertson

Abongwa

et al. 2016

International Journal for Parasitology: Drugs and Drug Resistan

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Soil-transmitted helminth infections in humans and livestock cause significant debility, reduced productivity and economic losses globally. There are a limited number of effective anthelmintic drugs available for treating helminths infections, and their frequent use has led to the development of resistance in many parasite species. There is an urgent need for novel therapeutic drugs for treating these parasites. We have chosen the ACR-16 nicotinic acetylcholine receptor of Ascaris suum (Asu-ACR-16), as a drug target and have developed three-dimensional models of this transmembrane protein receptor to facilitate the search for new bioactive compounds. Using the human α7 nAChR chimeras and Torpedo marmorata nAChR for homology modeling, we defined orthosteric and allosteric binding sites on the Asu-ACR-16 receptor for virtual screening. We identified four ligands that bind to sites on Asu-ACR-16 and tested their activity using electrophysiological recording from Asu-ACR-16 receptors expressed in Xenopus oocytes. The four ligands were acetylcholine inhibitors (SB-277011-A, IC50, 3.12 ± 1.29 μM; (+)-butaclamol Cl, IC50, 9.85 ± 2.37 μM; fmoc-1, IC50, 10.00 ± 1.38 μM; fmoc-2, IC50, 16.67 ± 1.95 μM) that behaved like negative allosteric modulators. Our work illustrates a structure-based in silico screening method for seeking anthelmintic hits, which can then be tested electrophysiologically for further characterization.

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Molecular Genetics of Torpedo Marmorata Acetylcholine Receptor

Cited by 7 publications

References 34 publications

Synthesis of an antigenic site of native acetylcholine receptor peptide 159-169 of Torpedo acetylcholine receptor α-chain

Synthesis of an antigenic site of native acetylcholine receptor peptide 159-169 of Torpedo acetylcholine receptor α-chain

Localization and synthesis of the acetylcholine-binding site in the α-chain of the Torpedo californica acetylcholine receptor

The Ascaris suum nicotinic receptor, ACR-16, as a drug target: Four novel negative allosteric modulators from virtual screening

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