S-glycosylation-based cysteine profiling reveals regulation of glycolysis by itaconate

• Published in: Nature chemical biology Vol. 15; no. 10; pp. 983 - 991
• Main Authors: Qin, Wei, Qin, Ke, Zhang, Yanling, Jia, Wentong, Chen, Ying, Cheng, Bo, Peng, Linghang, Chen, Nan, Liu, Yuan, Zhou, Wen, Wang, Yan-Ling, Chen, Xing, Wang, Chu
• Format: Journal Article
• Language: English
• Published: New York Nature Publishing Group US 01.10.2019; Nature Publishing Group
• Subjects: 631/92/1643; 631/92/458; 631/92/475; 631/92/96; Aldolase; Biochemical Engineering; Biochemistry; Biology; Bioorganic Chemistry; Cell Biology; Chemistry; Chemistry and Materials Science; Chemistry/Food Science; Cysteine; Datasets; Enzymes; Fructose; Fructose-bisphosphate aldolase; Glutathione; Glycolysis; Glycosylation; Immune response; Inflammation; Inflammatory response; Labeling; Laboratories; Macrophages; Mass spectrometry; Metabolism; Metabolites; Pathogens; Peptides; Proteins; Proteomes; Scientific imaging; Substrates
• ISSN: 1552-4450; 1552-4469; 1552-4469
• DOI: 10.1038/s41589-019-0323-5

Itaconate has been recently recognized as an anti-inflammatory metabolite involved in the pathogen–macrophage interface. Due to its weak electrophilicity, itaconate could modify cysteines of the protein KEAP1 and glutathione, which contribute to its anti-inflammatory effect. However, the substrates of itaconate modification in macrophages have not been systematically profiled, which largely impedes the understanding of its roles in immune responses. Here, we developed a specific thiol-reactive probe, 1-OH-Az, for quantitative chemoproteomic profiling of cysteine modifications by itaconate, and provided a global portrait of its proteome reactivity. We found that itaconate covalently modifies key glycolytic enzymes and impairs glycolytic flux mainly through inhibition of fructose-bisphosphate aldolase A (ALDOA). Moreover, itaconate attenuates the inflammatory response in stimulated macrophages by impairing the glycolysis. Our study provides a valuable resource of protein targets of itaconate in macrophages and establishes a negative-feedback link between glycolysis and itaconate, elucidating new functional insights for this anti-inflammatory metabolite. Using a thiol-reactive probe, chemoproteomic profiling of cysteine targets of itaconate reveals the covalent modification of glycolytic enzymes, impairing glycolytic flux and contributing to attenuation of the inflammatory response in macrophages.