Charting organellar importomes by quantitative mass spectrometry

Peikert, Christian D.; Mani, Jan; Morgenstern, Marcel; Käser, Sandro; Knapp, Bettina; Wenger, Christoph; Harsman, Anke Judith; Oeljeklaus, Silke; Schneider, André; Warscheid, Bettina (2017). Charting organellar importomes by quantitative mass spectrometry. Nature communications, 8, p. 15272. Nature Publishing Group 10.1038/ncomms15272

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Protein import into organelles is essential for all eukaryotes and facilitated by multi-protein translocation machineries. Analysing whether a protein is transported into an organelle is largely restricted to single constituents. This renders knowledge about imported proteins incomplete, limiting our understanding of organellar biogenesis and function. Here we introduce a method that enables charting an organelle's importome. The approach relies on inducible RNAi-mediated knockdown of an essential subunit of a translocase to impair import and quantitative mass spectrometry. To highlight its potential, we established the mitochondrial importome of Trypanosoma brucei, comprising 1,120 proteins including 331 new candidates. Furthermore, the method allows for the identification of proteins with dual or multiple locations and the substrates of distinct protein import pathways. We demonstrate the specificity and versatility of this ImportOmics method by targeting import factors in mitochondria and glycosomes, which demonstrates its potential for globally studying protein import and inventories of organelles.

Item Type:

Journal Article (Original Article)

Division/Institute:

08 Faculty of Science > Departement of Chemistry and Biochemistry

Graduate School:

Graduate School for Cellular and Biomedical Sciences (GCB)

UniBE Contributor:

Mani, Jan; Käser, Sandro; Wenger, Christoph; Harsman, Anke Judith and Schneider, André

Subjects:

500 Science > 570 Life sciences; biology
500 Science > 540 Chemistry

ISSN:

2041-1723

Publisher:

Nature Publishing Group

Language:

English

Submitter:

Christina Schüpbach

Date Deposited:

06 Jun 2017 16:06

Last Modified:

31 Aug 2017 08:59

Publisher DOI:

10.1038/ncomms15272

BORIS DOI:

10.7892/boris.100088

URI:

https://boris.unibe.ch/id/eprint/100088

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