Protein Painting Mass Spectrometry in the Discovery of Interaction Sites Within the Acetylcholine Binding Protein

Overview

Journal ACS Chem Neurosci

Publisher American Chemical Society

Specialty Neurology

Date 2024 May 28

PMID 38804618

Authors

Alexandru Graur

Amanda Haymond

Kyung Hyeon Lee

Franco Viscarra

Paul Russo

Alessandra Luchini

Mikell Paige

Isabel Bermudez-Diaz

Nadine Kabbani

Affiliations

Soon will be listed here.

Abstract

Nicotinic acetylcholine receptors (nAChRs) are a family of ligand-gated ion channel receptors that contribute to cognition, memory, and motor control in many organisms. The pharmacological targeting of these receptors, using small molecules or peptides, presents an important strategy for the development of drugs that can treat important human diseases, including neurodegenerative disorders. The acetylcholine binding protein (Ac-AChBP) is a structural surrogate of the nAChR with high homology to the extracellular ligand binding domain of homopentameric nAChRs. In this study, we optimized protein-painting-based mass spectrometry to identify regions of interaction between the Ac-AChBP and several nAChR ligands. Using molecular dyes that adhere to the surface of a solubilized Ac-AChBP complex, we identified amino acid residues that constitute a contact site within the Ac-AChBP for α-bungarotoxin, choline, nicotine, and amyloid-β 1-42. By integrating innovation in protein painting mass spectrometry with computational structural modeling, we present a new experimental tool for analyzing protein interactions of the nAChR.

Citing Articles

Using Protein Painting Mass Spectrometry to Define Ligand Receptor Interaction Sites for Acetylcholine Binding Protein.

Graur A, Erickson N, Kabbani N Bio Protoc. 2025; 15(2):e5163.

PMID: 39872720 PMC: 11769746. DOI: 10.21769/BioProtoc.5163.

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