Probing Ion Channel Neighborhoods Using Proximity Proteomics

Overview

Journal Methods Enzymol

Specialty Biochemistry

Date 2021 Jun 14

PMID 34120710

Citations 2

Authors

Gabriel Redel-Traub

Guoxia Liu

Steven O Marx

Jared Kushner

Affiliations

Soon will be listed here.

Abstract

Protein-protein interactions are critically important for cellular functions, including regulation of ion channels. Ion channels are typically part of large macromolecular complexes that impact their function. These complexes have traditionally been elucidated via standard biochemical techniques including immunoprecipitation, pull-down assays and mass spectrometry. Recently, several methods have been developed to provide a more complete depiction of the microenvironment or "neighborhood" of proteins of interest. These new methods, which fall broadly under the category of proximity-dependent labeling techniques, aim to overcome the limitations imposed by antibody-based techniques and mass spectrometry. In this chapter, we describe the use of proximity labeling to elucidate the cardiac Ca1.2 macromolecular complex under basal conditions and after β-adrenergic stimulation. Using these methodologies, we have identified the mechanism underlying adrenergic stimulation of the Ca current in the heart.

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