%0 Journal Article %1 peikertChartingOrganellarImportomes2017a %A Peikert, Christian D. %A Mani, Jan %A Morgenstern, Marcel %A Käser, Sandro %A Knapp, Bettina %A Wenger, Christoph %A Harsman, Anke %A Oeljeklaus, Silke %A Schneider, André %A Warscheid, Bettina %C England %D 2017 %J Nature communications %K Acyl-tRNA Amino Knockdown Membranes/metabolism,Mitochondrial Proteins/*metabolism,Protein Proteins/*metabolism,Substrate Specificity,to_read,Trypanosoma Spectrometry/*methods,Microbodies/metabolism,Mitochondria/*metabolism,Mitochondrial Synthetases/metabolism,Gene Techniques,Mass Transport,Proteome/*metabolism,Protozoan brucei brucei/*metabolism %P 15272 %R 10.1038/ncomms15272 %T Charting Organellar Importomes by Quantitative Mass Spectrometry. %V 8 %X 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.

@article{peikertChartingOrganellarImportomes2017a, abstract = {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.}, added-at = {2024-05-17T13:01:35.000+0200}, address = {England}, author = {Peikert, Christian D. and Mani, Jan and Morgenstern, Marcel and K{\"a}ser, Sandro and Knapp, Bettina and Wenger, Christoph and Harsman, Anke and Oeljeklaus, Silke and Schneider, Andr{\'e} and Warscheid, Bettina}, biburl = {https://www.bibsonomy.org/bibtex/2fc769f7a96aa552f9ae37a03fcca85b3/warscheidlab}, doi = {10.1038/ncomms15272}, interhash = {fba8e49674c6acd97d65b7f41c4e6018}, intrahash = {fc769f7a96aa552f9ae37a03fcca85b3}, issn = {2041-1723}, journal = {Nature communications}, keywords = {Acyl-tRNA Amino Knockdown Membranes/metabolism,Mitochondrial Proteins/*metabolism,Protein Proteins/*metabolism,Substrate Specificity,to_read,Trypanosoma Spectrometry/*methods,Microbodies/metabolism,Mitochondria/*metabolism,Mitochondrial Synthetases/metabolism,Gene Techniques,Mass Transport,Proteome/*metabolism,Protozoan brucei brucei/*metabolism}, langid = {english}, month = may, pages = 15272, pmcid = {PMC5436138}, pmid = {28485388}, timestamp = {2024-05-17T13:01:35.000+0200}, title = {Charting Organellar Importomes by Quantitative Mass Spectrometry.}, volume = 8, year = 2017 }