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Proteomic mapping in live Drosophila tissues using an engineered ascorbate peroxidase

Proceedings of the National Academy of Sciences - PNAS, 2015-09, Vol.112 (39), p.12093-12098 [Peer Reviewed Journal]

Copyright National Academy of Sciences Sep 29, 2015 ;ISSN: 0027-8424 ;EISSN: 1091-6490 ;DOI: 10.1073/pnas.1515623112 ;PMID: 26362788

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Title:
Proteomic mapping in live Drosophila tissues using an engineered ascorbate peroxidase
Author: Chen, Chiao-Lin ; Hu, Yanhui ; Udeshi, Namrata D ; Lau, Thomas Y ; Wirtz-Peitz, Frederik ; He, Li ; Ting, Alice Y ; Carr, Steven A ; Perrimon, Norbert
Subjects: Animals ; Ascorbate Peroxidases - genetics ; Biological Sciences ; Computational Biology ; Databases, Protein ; Drosophila ; Drosophila - genetics ; Drosophila - metabolism ; Enzymes ; Genetic Engineering - methods ; Insects ; Mitochondria ; Proteomics ; Proteomics - methods ; Staining and Labeling - methods
Is Part Of: Proceedings of the National Academy of Sciences - PNAS, 2015-09, Vol.112 (39), p.12093-12098
Description: 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.
Publisher: United States: National Acad Sciences
Language: English
Identifier: ISSN: 0027-8424
EISSN: 1091-6490
DOI: 10.1073/pnas.1515623112
PMID: 26362788
Source: Geneva Foundation Free Medical Journals at publisher websites
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Proteomic mapping in live Drosophila tissues using an engineered ascorbate peroxidase

Copyright National Academy of Sciences Sep 29, 2015 ;ISSN: 0027-8424 ;EISSN: 1091-6490 ;DOI: 10.1073/pnas.1515623112 ;PMID: 26362788

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