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Charting organellar importomes by quantitative mass spectrometry.

  • Peikert CD Faculty of Biology, Department of Biochemistry and Functional Proteomics, Institute of Biology II, University of Freiburg, Schänzlestrasse 1, 79104 Freiburg, Germany.
  • Mani J Department of Chemistry and Biochemistry, University of Bern, Freiestrasse 3, CH-3012 Bern, Switzerland.
  • Morgenstern M Faculty of Biology, Department of Biochemistry and Functional Proteomics, Institute of Biology II, University of Freiburg, Schänzlestrasse 1, 79104 Freiburg, Germany.
  • Käser S Department of Chemistry and Biochemistry, University of Bern, Freiestrasse 3, CH-3012 Bern, Switzerland.
  • Knapp B Faculty of Biology, Department of Biochemistry and Functional Proteomics, Institute of Biology II, University of Freiburg, Schänzlestrasse 1, 79104 Freiburg, Germany.
  • Wenger C Department of Chemistry and Biochemistry, University of Bern, Freiestrasse 3, CH-3012 Bern, Switzerland.
  • Harsman A Department of Chemistry and Biochemistry, University of Bern, Freiestrasse 3, CH-3012 Bern, Switzerland.
  • Oeljeklaus S Faculty of Biology, Department of Biochemistry and Functional Proteomics, Institute of Biology II, University of Freiburg, Schänzlestrasse 1, 79104 Freiburg, Germany.
  • Schneider A Department of Chemistry and Biochemistry, University of Bern, Freiestrasse 3, CH-3012 Bern, Switzerland.
  • Warscheid B Faculty of Biology, Department of Biochemistry and Functional Proteomics, Institute of Biology II, University of Freiburg, Schänzlestrasse 1, 79104 Freiburg, Germany.
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  • 2017-05-10
Published in:
  • Nature communications. - 2017
Amino Acyl-tRNA Synthetases
Gene Knockdown Techniques
Mass Spectrometry
Microbodies
Mitochondria
Mitochondrial Membranes
Mitochondrial Proteins
Protein Transport
Proteome
Protozoan Proteins
Substrate Specificity
Trypanosoma brucei brucei
English 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.
Language
  • English
Open access status
gold
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Persistent URL
https://sonar.ch/global/documents/178290
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