1. Mapping New Residents of the Mitochondrial Nucleoid
Laura Trinkle-Mulcahy 
Cell Chemical Biology 
Volume 24, Issue 3, Pages (March 2017)
DOI: /j.chembiol
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2. Figure 1
Design of Quantitative Ratiometric APEX2-Based Mapping of the Mitochondrial Nucleoid Proteome
The strategy utilized to highlight nucleoid-specific proteins was based on stable expression of the nucleoid protein Twinkle fused to APEX2 in HEK293 cells and the addition of biotin-phenol and H2O2 to catalyze biotinylation of neighboring proteins. Control conditions that were included to flag non-specific hits omitted either H2O2 treatment or expression of Twinkle-APEX2. An additional control that was added to highlight nucleoid-specific proteins above the background of other mitochondrial proteins was HEK293 cells stably expressing mito-APEX2, which targets the peroxidase throughout the entire mitochondrial matrix. Cells were harvested for lysis and extraction/capture of biotinylated proteins on streptavidin-coated magnetic beads. Trypsin digestion, tandem mass tag (TMT) labeling and LC-MS/MS were used to identify proteins and quantify their relative abundance in the three conditions.
Cell Chemical Biology  , DOI: ( /j.chembiol )
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