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Protein Modification by Endogenously Generated Lipid Electrophiles: Mitochondria as the Source and Target
journal contribution
posted on 2017-06-14, 00:00 authored by William
N. Beavers, Kristie L. Rose, James J. Galligan, Michelle M. Mitchener, Carol A. Rouzer, Keri A. Tallman, Connor R. Lamberson, Xiaojing Wang, Salisha Hill, Pavlina T. Ivanova, H. Alex Brown, Bing Zhang, Ned A. Porter, Lawrence J. MarnettDetermining
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.