title: Chemical crosslinkers enhance detection of receptor interactomes.
abstract: Receptor function is dependent on interaction with various intracellular proteins that ensure the localization and signaling of the receptor. While a number of approaches have been optimized for the isolation, purification, and proteomic characterization of receptor-protein interaction networks (interactomes) in cells, the capture of receptor interactomes and their dynamic properties remains a challenge. In particular, the study of interactome components that bind to the receptor with low affinity or can rapidly dissociate from the macromolecular complex is difficult. Here we describe how chemical crosslinking (CC) can aid in the isolation and proteomic analysis of receptor-protein interactions. The addition of CC to standard affinity purification and mass spectrometry protocols boosts the power of protein capture within the proteomic assay and enables the identification of specific binding partners under various cellular and receptor states. The utility of CC in receptor interactome studies is highlighted for the nicotinic acetylcholine receptor as well as several other receptor types. A better understanding of receptors and their interactions with proteins spearheads molecular biology, informs an integral part of bench medicine which helps in drug development, drug action, and understanding the pathophysiology of disease.
authors.affiliation: Department of Molecular Neuroscience, Krasnow Institute for Advanced Study, George Mason University Fairfax, VA, USA.
authors.forename: Brian A
authors.initials: BA
authors.lastname: Corgiat
authors.affiliation: Department of Molecular Neuroscience, Krasnow Institute for Advanced Study, George Mason University Fairfax, VA, USA.
authors.forename: Jacob C
authors.initials: JC
authors.lastname: Nordman
authors.affiliation: Department of Molecular Neuroscience, Krasnow Institute for Advanced Study, George Mason University Fairfax, VA, USA.
authors.forename: Nadine
authors.initials: N
authors.lastname: Kabbani
keywords: chemical crosslinking; interactome; mass spectrometry; nicotinic acetylcholine receptor; protein–protein interaction; signaling network