Characterisation of the plasma membrane subproteome of bloodstream form Trypanosoma brucei

Daniel J. Bridges, Andrew R Pitt, Orla Hanrahan, Kiva Brennan, H. Paul Voorheis, Pawel Herzyk, Harry P. de Koning, Richard J S Burchmore

Research output: Contribution to journalArticle

Abstract

Proteome analysis by conventional approaches is biased against hydrophobic membrane proteins, many of which are also of low abundance. We have isolated plasma membrane sheets from bloodstream forms of Trypanosoma brucei by subcellular fractionation, and then applied a battery of complementary protein separation and identification techniques to identify a large number of proteins in this fraction. The results of these analyses have been combined to generate a subproteome for the pellicular plasma membrane of bloodstream forms of T. brucei as well as a separate subproteome for the pellicular cytoskeleton. In parallel, we have used in silico approaches to predict the relative abundance of proteins potentially expressed by bloodstream form trypanosomes, and to identify likely polytopic membrane proteins, providing quality control for the experimentally defined plasma membrane subproteome. We show that the application of multiple high-resolution proteomic techniques to an enriched organelle fraction is a valuable approach for the characterisation of relatively intractable membrane proteomes. We present here the most complete analysis of a protozoan plasma membrane proteome to date and show the presence of a large number of integral membrane proteins, including 11 nucleoside/nucleobase transporters, 15 ion pumps and channels and a large number of adenylate cyclases hitherto listed as putative proteins.
LanguageEnglish
Pages83-99
Number of pages17
JournalProteomics
Volume8
Issue number1
Early online date19 Dec 2007
DOIs
Publication statusPublished - Jan 2008

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Trypanosoma brucei brucei
Cell membranes
Proteome
Cell Membrane
Membrane Proteins
Proteins
Ion Pumps
Nucleoside Transport Proteins
Trypanosomiasis
Fractionation
Cytoskeleton
Ion Channels
Nucleosides
Adenylyl Cyclases
Quality Control
Organelles
Computer Simulation
Proteomics
Quality control
Membranes

Keywords

  • animals
  • cell membrane
  • liquid chromatography
  • codon
  • female
  • membrane proteins
  • protein array analysis
  • proteome
  • protozoan proteins
  • rats
  • Wistar rats
  • spectrometry
  • tandem mass spectrometry
  • trypanosoma brucei brucei
  • mass
  • electrospray ionization

Cite this

Bridges, D. J., Pitt, A. R., Hanrahan, O., Brennan, K., Voorheis, H. P., Herzyk, P., ... Burchmore, R. J. S. (2008). Characterisation of the plasma membrane subproteome of bloodstream form Trypanosoma brucei. Proteomics, 8(1), 83-99. https://doi.org/10.1002/pmic.200700607
Bridges, Daniel J. ; Pitt, Andrew R ; Hanrahan, Orla ; Brennan, Kiva ; Voorheis, H. Paul ; Herzyk, Pawel ; de Koning, Harry P. ; Burchmore, Richard J S. / Characterisation of the plasma membrane subproteome of bloodstream form Trypanosoma brucei. In: Proteomics. 2008 ; Vol. 8, No. 1. pp. 83-99.
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Bridges, DJ, Pitt, AR, Hanrahan, O, Brennan, K, Voorheis, HP, Herzyk, P, de Koning, HP & Burchmore, RJS 2008, 'Characterisation of the plasma membrane subproteome of bloodstream form Trypanosoma brucei' Proteomics, vol. 8, no. 1, pp. 83-99. https://doi.org/10.1002/pmic.200700607

Characterisation of the plasma membrane subproteome of bloodstream form Trypanosoma brucei. / Bridges, Daniel J.; Pitt, Andrew R; Hanrahan, Orla; Brennan, Kiva; Voorheis, H. Paul; Herzyk, Pawel; de Koning, Harry P.; Burchmore, Richard J S.

In: Proteomics, Vol. 8, No. 1, 01.2008, p. 83-99.

Research output: Contribution to journalArticle

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