Proteomic mapping in live Drosophila tissues using an engineered ascorbate peroxidase
Name
Chen-2015-Proteomic mapping in.pdf
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1.47 MB
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Checksum (MD5)
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Author(s)
Chen, Chiao-Lin
Hu, Yanhui
Udeshi, Namrata D.
Lau, Thomas Y.
Wirtz-Peitz, Frederik
He, Li
Ting, Alice Y.
Carr, Steven A.
Perrimon, Norbert
Date Issued
September 2015
Journal
Proceedings of the National Academy of Sciences
Publisher
National Academy of Sciences (U.S.)
Citation
Chen, Chiao-Lin, Yanhui Hu, Namrata D. Udeshi, Thomas Y. Lau, Frederik Wirtz-Peitz, Li He, Alice Y. Ting, Steven A. Carr, and Norbert Perrimon. “ Proteomic Mapping in Live Drosophila Tissues Using an Engineered Ascorbate Peroxidase .” Proc Natl Acad Sci USA 112, no. 39 (September 11, 2015): 12093–12098.
Version
Final published version
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.
MIT Department
Massachusetts Institute of Technology. Department of Chemistry
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Article is made available in accordance with the publisher's policy and may be subject to US copyright law. Please refer to the publisher's site for terms of use.
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DOI of Published Version
https://doi.org/10.1073/pnas.1515623112
