You are here

Proc Natl Acad Sci U S A DOI:10.1073/pnas.1515623112

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

Publication TypeJournal Article
Year of Publication2015
AuthorsChen, C-L, Hu, Y, Udeshi, ND, Lau, TY, Wirtz-Peitz, F, He, L, Ting, AY, Carr, SA, Perrimon, N
JournalProc Natl Acad Sci U S A
Volume112
Issue39
Pages12093-8
Date Published2015 Sep 29
ISSN1091-6490
KeywordsAnimals, Ascorbate Peroxidases, Computational Biology, Databases, Protein, Drosophila, Genetic Engineering, Proteomics, Staining and Labeling
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 that provides an inventory of Drosophila mitochondrial proteins with subcompartmental annotation. Altogether, APEX labeling in live Drosophila tissues provides an opportunity to characterize the organelle proteome of specific cell types in different physiological conditions.

URLhttp://www.pnas.org/cgi/pmidlookup?view=long&pmid=26362788
DOI10.1073/pnas.1515623112
Pubmed

http://www.ncbi.nlm.nih.gov/pubmed/26362788?dopt=Abstract

Alternate JournalProc. Natl. Acad. Sci. U.S.A.
PubMed ID26362788
PubMed Central IDPMC4593093
Grant ListR21 ES025615 / ES / NIEHS NIH HHS / United States
P01 CA120964 / CA / NCI NIH HHS / United States
R01 DK088718 / DK / NIDDK NIH HHS / United States
/ / Howard Hughes Medical Institute / United States
R01-DK088718 / DK / NIDDK NIH HHS / United States
P01-CA120964 / CA / NCI NIH HHS / United States