ACS Chemical Biology
ACS Chemical Biology
5 years ago

Protein Modification by Endogenously Generated Lipid Electrophiles: Mitochondria as the Source and Target

Protein Modification by Endogenously Generated Lipid Electrophiles: Mitochondria as the Source and Target
William N. Beavers, Bing Zhang, Xiaojing Wang, Salisha Hill, James J. Galligan, Ned A. Porter, Michelle M. Mitchener, Lawrence J. Marnett, H. Alex Brown, Kristie L. Rose, Keri A. Tallman, Carol A. Rouzer, Pavlina T. Ivanova, Connor R. Lamberson
Determining the impact of lipid electrophile-mediated protein damage that occurs during oxidative stress requires a comprehensive analysis of electrophile targets adducted under pathophysiological conditions. Incorporation of ω-alkynyl linoleic acid into the phospholipids of macrophages prior to activation by Kdo2-lipid A, followed by protein extraction, click chemistry, and streptavidin affinity capture, enabled a systems-level survey of proteins adducted by lipid electrophiles generated endogenously during the inflammatory response. Results revealed a dramatic enrichment for membrane and mitochondrial proteins as targets for adduction. A marked decrease in adduction in the presence of MitoTEMPO demonstrated a primary role for mitochondrial superoxide in electrophile generation and indicated an important role for mitochondria as both a source and target of lipid electrophiles, a finding that has not been revealed by prior studies using exogenously provided electrophiles.

Publisher URL: http://dx.doi.org/10.1021/acschembio.7b00480

DOI: 10.1021/acschembio.7b00480

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