Architecture Mapping of the Inner Mitochondrial Membrane Proteome by Chemical Tools in Live Cells

• DOI: 10.1021/jacs.6b10418
• Publication Date: 2017-02-03T16:00:42Z
• Authors: Song-Yi Lee; Myeong-Gyun Kang; Sanghee Shin; Chulhwan Kwak; Taejoon Kwon; Jeong Kon Seo; Jong-Seo Kim; Hyun-Woo Rhee
• Keywords: Proteome; Membrane topology

The inner mitochondrial membrane (IMM) proteome plays a central role in maintaining physiology and cellular metabolism. Various important biochemical reactions such as oxidative phosphorylation, metabolite production, biogenesis are conducted by the IMM proteome, mitochondria-targeted therapeutics have been developed for proteins, which is deeply related various human metabolic diseases including cancer neurodegenerative diseases. However, topology of remains largely unclear because lack methods to evaluate it live cells high-throughput manner. In this article, we reveal vivo topological direction 135 using an situ-generated radical probe with genetically targeted peroxidase (APEX). Owing short lifetime phenoxyl radicals generated situ submitochondrial APEX impermeability small molecules, solvent-exposed tyrosine residues both matrix intermembrane space (IMS) sides proteins were exclusively labeled Matrix-APEX IMS-APEX, respectively identified mass spectrometry. From analysis, confirmed 58 protein topologies could determine 77 whose at has not fully characterized. We also found several (e.g., LETM1 OXA1) information should be revised on basis our results. Overall, identification structural inner-membrane can provide valuable insights architecture connectome cells.