Issue 20, 2022

A proximity labeling method for protein–protein interactions on cell membrane

Abstract

Antibodies targeting specific antigens are widely utilized in biological research to investigate protein interactions or to quantify target antigens. Here, we introduce antigen–antibody proximity labeling (AAPL), a novel method to map the antigen interaction sites as well as interactors of antibody-targeted proteins. As a proof of concept, AAPL was demonstrated using sodium/potassium transporting ATPase (ATP1A1) and epidermal growth factor receptor 2 (ERBB2)-specific antibodies that were modified with an Fe(III) catalytic probe. Once bound to their target proteins, Fe(III)-induced catalytic oxidation occurred in proximity of the antigen's epitope. Oxidative proteomic analysis was then used to determine the degree of oxidation, the site of oxidation within the targeted antigen, and the interacting proteins that were in close proximity to the targeted antigen. An AAPL score was generated for each protein yielding the specificity of the oxidation and proximity of the interacting protein to the target antigen. As a final demonstration of its utility, the AAPL approach was applied to map the interactors of liver–intestine-cadherin (CDH17) in colon cancer cells.

Graphical abstract: A proximity labeling method for protein–protein interactions on cell membrane

Supplementary files

Article information

Article type
Edge Article
Submitted
10 Dec 2021
Accepted
29 Apr 2022
First published
30 Apr 2022
This article is Open Access

All publication charges for this article have been paid for by the Royal Society of Chemistry
Creative Commons BY license

Chem. Sci., 2022,13, 6028-6038

A proximity labeling method for protein–protein interactions on cell membrane

Q. Li, Y. Xie, R. Rice, E. Maverakis and C. B. Lebrilla, Chem. Sci., 2022, 13, 6028 DOI: 10.1039/D1SC06898A

This article is licensed under a Creative Commons Attribution 3.0 Unported Licence. You can use material from this article in other publications without requesting further permissions from the RSC, provided that the correct acknowledgement is given.

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