Proteomic analysis of the cell cycle of procylic form Trypanosoma brucei

Thomas W. M. Crozier, Michele Tinti, Richard J. Wheeler, Tony Ly, Michael A. J. Ferguson, Angus I. Lamond (Lead / Corresponding author)

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Abstract

We describe a single-step centrifugal elutriation method to produce synchronous G1-phase procyclic trypanosomes at a scale amenable for proteomic analysis of the cell cycle. Using ten-plex tandem mass tag (TMT) labelling and mass spectrometry (MS)-based proteomics technology, the expression levels of 5,325 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.

Original languageEnglish
Pages (from-to)1184-1195
Number of pages12
JournalMolecular & Cellular Proteomics
Volume17
Issue number6
Early online date19 Mar 2018
DOIs
Publication statusPublished - 1 Jun 2018

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Keywords

  • Journal article
  • Trypanosoma
  • Procyclic
  • Cell cycle
  • Proteomics
  • Tandem mass tagging
  • Cell cycle*
  • Quantification
  • Mitosis
  • Parasite
  • Molecular biology*
  • Infectious disease
  • Mass Spectrometry
  • procyclic
  • Cell biology*
  • Cell division*
  • Gene Expression*

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