Proteomic Analysis of the Cell Cycle of Procylic Form Trypanosoma brucei .
Autor: | Crozier TWM; From the ‡Wellcome Centre for Anti-Infectives Research, School of Life Sciences, University of Dundee, Dundee, DD1 5EH, UK.; §Centre for Gene Regulation and Expression, School of Life Sciences, University of Dundee, Dundee, DD1 5EH, UK., Tinti M; From the ‡Wellcome Centre for Anti-Infectives Research, School of Life Sciences, University of Dundee, Dundee, DD1 5EH, UK., Wheeler RJ; ‖Sir William Dunn School of Pathology, University of Oxford, Oxford, OX1 3RE, UK., Ly T; §Centre for Gene Regulation and Expression, School of Life Sciences, University of Dundee, Dundee, DD1 5EH, UK., Ferguson MAJ; From the ‡Wellcome Centre for Anti-Infectives Research, School of Life Sciences, University of Dundee, Dundee, DD1 5EH, UK; a.i.lamond@dundee.ac.uk m.a.j.ferguson@dundee.ac.uk., Lamond AI; §Centre for Gene Regulation and Expression, School of Life Sciences, University of Dundee, Dundee, DD1 5EH, UK; a.i.lamond@dundee.ac.uk m.a.j.ferguson@dundee.ac.uk. |
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Jazyk: | angličtina |
Zdroj: | Molecular & cellular proteomics : MCP [Mol Cell Proteomics] 2018 Jun; Vol. 17 (6), pp. 1184-1195. Date of Electronic Publication: 2018 Mar 19. |
DOI: | 10.1074/mcp.RA118.000650 |
Abstrakt: | We describe a single-step centrifugal elutriation method to produce synchronous Gap1 (G1)-phase procyclic trypanosomes at a scale amenable for proteomic analysis of the cell cycle. Using ten-plex tandem mass tag (TMT) labeling and mass spectrometry (MS)-based proteomics technology, the expression levels of 5325 proteins were quantified across the cell cycle in this parasite. Of these, 384 proteins were classified as cell-cycle regulated and subdivided into nine clusters with distinct temporal regulation. These groups included many known cell cycle regulators in trypanosomes, which validates the approach. In addition, we identify 40 novel cell cycle regulated proteins that are essential for trypanosome survival and thus represent potential future drug targets for the prevention of trypanosomiasis. Through cross-comparison to the TrypTag endogenous tagging microscopy database, we were able to validate the cell-cycle regulated patterns of expression for many of the proteins of unknown function detected in our proteomic analysis. A convenient interface to access and interrogate these data is also presented, providing a useful resource for the scientific community. Data are available via ProteomeXchange with identifier PXD008741 (https://www.ebi.ac.uk/pride/archive/). (© 2018 by The American Society for Biochemistry and Molecular Biology, Inc.) |
Databáze: | MEDLINE |
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