Activity-based protein profiling of skeletal muscle mitochondria with ATP-ABP.

<p>(A) Probe structure of <b>ATP-ABP</b>, and reactivity with ATPases and ATP-binding proteins. The ε-amino group of lysine reacts with the mixed anhydride of <b>ATP-ABP</b> to form a stable amide bond on the protein to a six-carbon moiety containing the click chemistry (CC) compatible alkyne. Subsequent CC reaction to biotin-azide permits enrichment of <b>ATP-ABP</b>-labeled proteins. (B) Sample preparation of <b>ATP-ABP</b> labeled skeletal muscle mitochondrial proteins. Mitochondria were extracted from skeletal muscle tissue from individual mice, labeled with the probe (pink square), captured on streptavidin agarose resin (yellow circle), digested on-resin with trypsin, and the subsequent peptides were measured by LC-MS/MS.