ArticleThe Journal of biological chemistry2002
Tryptophan fluorescence reveals conformational changes in the acetylcholine binding protein.
Article in The Journal of biological chemistry, 2002. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 43 papers.
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Who cites it
43 citing papers in PubMed, 99 citations in OpenAlex.
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- Interactions of Nereistoxin and Its Analogs with Vertebrate Nicotinic Acetylcholine Receptors and Molluscan ACh Binding Proteins.Marine drugs · 2022Article
- Snake Toxins Labeled by Green Fluorescent Protein or Its Synthetic Chromophore are New Probes for Nicotinic acetylcholine Receptors.Frontiers in molecular biosciences · 2021Article
- Molecular cloning and biochemical characterization of the phospholipid scramblase SCRM-1 from Caenorhabditis elegans.European biophysics journal : EBJ · 2020Article
- Structure-Function of Neuronal Nicotinic Acetylcholine Receptor Inhibitors Derived From Natural Toxins.Frontiers in neuroscience · 2020Review
- Escherichia coli Protein Expression System for Acetylcholine Binding Proteins (AChBPs).PloS one · 2016Article
- Marine Macrocyclic Imines, Pinnatoxins A and G: Structural Determinants and Functional Properties to Distinguish Neuronal α7 from Muscle α1(2)βγδ nAChRs.Structure (London, England : 1993) · 2015Article
- EPR Studies of Gating Mechanisms in Ion Channels.Methods in enzymology · 2015Review
- Iron inhibits Escherichia coli topoisomerase I activity by targeting the first two zinc-binding sites in the C-terminal domain.Protein science : a publication of the Protein Society · 2014Article
- Structural basis for cooperative interactions of substituted 2-aminopyrimidines with the acetylcholine binding protein.Proceedings of the National Academy of Sciences of the United States of America · 2014Article
- Efficient expression of acetylcholine-binding protein from Aplysia californica in Bac-to-Bac system.BioMed research international · 2014Article
- Article
- Development of a solid-phase receptor-based assay for the detection of cyclic imines using a microsphere-flow cytometry system.Analytical chemistry · 2013Article
- Assembly of a π-π stack of ligands in the binding site of an acetylcholine-binding protein.Nature communications · 2013Article
- End-plate acetylcholine receptor: structure, mechanism, pharmacology, and disease.Physiological reviews · 2012Review
- Desensitization mechanism in prokaryotic ligand-gated ion channel.The Journal of biological chemistry · 2012Article
- Intersubunit bridge formation governs agonist efficacy at nicotinic acetylcholine α4β2 receptors: unique role of halogen bonding revealed.The Journal of biological chemistry · 2012Article
- Creating an α7 nicotinic acetylcholine recognition domain from the acetylcholine-binding protein: crystallographic and ligand selectivity analyses.The Journal of biological chemistry · 2011Article
- Neurotransmitter/sodium symporter orthologue LeuT has a single high-affinity substrate site.Nature · 2010Article
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Authors and funding
7 authors at 1 institution in 1 country.
Funding
Abstract
The recent characterization of an acetylcholine binding protein (AChBP) from the fresh water snail, Lymnaea stagnalis, shows it to be a structural homolog of the extracellular domain of the nicotinic acetylcholine receptor (nAChR). To ascertain whether the AChBP exhibits the recognition properties and functional states of the nAChR, we have expressed the protein in milligram quantities from a synthetic cDNA transfected into human embryonic kidney (HEK) cells. The protein secreted into the medium shows a pentameric rosette structure with ligand stoichiometry approximating five sites per pentamer. Surprisingly, binding of acetylcholine, selective agonists, and antagonists ranging from small alkaloids to larger peptides results in substantial quenching of the intrinsic tryptophan fluorescence. Using stopped-flow techniques, we demonstrate rapid rates of association and dissociation of agonists and slow rates for the alpha-neurotoxins. Since agonist binding occurs in millisecond time frames, and the alpha-neurotoxins may induce a distinct conformational state for the AChBP-toxin complex, the snail protein shows many of the properties expected for receptor recognition of interacting ligands. Thus, the marked tryptophan quenching not only documents the importance of aromatic residues in ligand recognition, but establishes that the AChBP will be a useful functional as well as structural surrogate of the nicotinic receptor.
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