Proteomic mapping of the human mitochondrial intermembrane space in live cells via ratiometric APEX tagging.

ABSTRACT: Obtaining complete protein inventories for subcellular regions is a challenge that often limits our understanding of cellular function, especially for regions that are impossible to purify and are therefore inaccessible to traditional proteomic analysis. We recently developed a method to map proteomes in living cells with an engineered peroxidase (APEX) that bypasses the need for organellar purification when applied to membrane-bound compartments; however, it was insufficiently specific when applied to unbounded regions that allow APEX-generated radicals to escape. Here, we combine APEX technology with a SILAC-based ratiometric tagging strategy to substantially reduce unwanted background and achieve nanometer spatial resolution. This is applied to map the proteome of the mitochondrial inte

SUBMITTER: Hung V 

PROVIDER: S-EPMC4743503 | biostudies-literature | 2014 Jul

REPOSITORIES: biostudies-literature