Proteomic mapping in live Drosophila tissues using an engineered ascorbate peroxidase.

ABSTRACT: Characterization of the proteome of organelles and subcellular domains is essential for understanding cellular organization and identifying protein complexes as well as networks of protein interactions. We established a proteomic mapping platform in live Drosophila tissues using an engineered ascorbate peroxidase (APEX). Upon activation, the APEX enzyme catalyzes the biotinylation of neighboring endogenous proteins that can then be isolated and identified by mass spectrometry. We demonstrate that APEX labeling functions effectively in multiple fly tissues for different subcellular compartments and maps the mitochondrial matrix proteome of Drosophila muscle to demonstrate the power of APEX for characterizing subcellular proteomes in live cells. Further, we generate "MitoMax," a database tha

SUBMITTER: Chen CL 

PROVIDER: S-EPMC4593093 | biostudies-literature | 2015 Sep

REPOSITORIES: biostudies-literature