Issue 30, 2025

Thermal proteome profiling of itaconate interactome in macrophages

Abstract

Itaconate (ITA) is an upregulated immunometabolite in macrophages during pathogen infection. It is known to influence oxidation stress, cellular metabolism, programmed cell death and many other biological processes to regulate the immune response via interaction with proteins. Previous studies capture covalently ITA-modified proteins by activity-based proteome profiling with bioorthogonal chemical probes; however, how itaconate interacts non-covalently with other proteins at the proteome level remains unexplored. Here we applied thermal proteome profiling (TPP) to globally identify a large number of ITA-interacting proteins in macrophage proteomes. Among these targets, we verified mitochondrial branched-chain aminotransferase (BCAT2) as a novel non-covalent binding target of itaconate via biochemical and structural experiments. The binding of itaconate could inhibit transamination activity of BCAT and regulate the metabolism of branched-chain amino acids (BCAAs) in lipopolysaccharide (LPS)-activated inflammatory macrophages. This study offers a valuable resource that helps decipher novel and comprehensive functions of ITA in macrophages during the immune response and other related biological processes.

Graphical abstract: Thermal proteome profiling of itaconate interactome in macrophages

Supplementary files

Article information

Article type
Edge Article
Submitted
29 Mar 2025
Accepted
22 Jun 2025
First published
02 Jul 2025
This article is Open Access

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

Chem. Sci., 2025,16, 13838-13846

Thermal proteome profiling of itaconate interactome in macrophages

Y. Meng, T. Wei, C. Zhang, A. Yu, Y. Liu, J. Xiao and C. Wang, Chem. Sci., 2025, 16, 13838 DOI: 10.1039/D5SC02378E

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