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Eukaryotic Cell, August 2007, p. 1450-1463, Vol. 6, No. 8
1535-9778/07/$08.00+0     doi:10.1128/EC.00175-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Sugar Nucleotide Pools of Trypanosoma brucei, Trypanosoma cruzi, and Leishmania major{triangledown} ,{dagger}

Daniel C. Turnock and Michael A. J. Ferguson*

Division of Biological Chemistry and Drug Discovery, Wellcome Trust Biocentre, College of Life Sciences, University of Dundee, Dundee DD1 5EH, Scotland, United Kingdom

Received 16 May 2007/ Accepted 29 May 2007

The cell surface glycoconjugates of trypanosomatid parasites are intimately involved in parasite survival, infectivity, and virulence in their insect vectors and mammalian hosts. Although there is a considerable body of work describing their structure, biosynthesis, and function, little is known about the sugar nucleotide pools that fuel their biosynthesis. In order to identify and quantify parasite sugar nucleotides, we developed an analytical method based on liquid chromatography-electrospray ionization-tandem mass spectrometry using multiple reaction monitoring. This method was applied to the bloodstream and procyclic forms of Trypanosoma brucei, the epimastigote form of T. cruzi, and the promastigote form of Leishmania major. Five sugar nucleotides, GDP-{alpha}-D-mannose, UDP-{alpha}-D-N-acetylglucosamine, UDP-{alpha}-D-glucose, UDP-{alpha}-galactopyranose, and GDP-ß-L-fucose, were common to all three species; one, UDP-{alpha}-D-galactofuranose, was common to T. cruzi and L. major; three, UDP-ß-L-rhamnopyranose, UDP-{alpha}-D-xylose, and UDP-{alpha}-D-glucuronic acid, were found only in T. cruzi; and one, GDP-{alpha}-D-arabinopyranose, was found only in L. major. The estimated demands for each monosaccharide suggest that sugar nucleotide pools are turned over at very different rates, from seconds to hours. The sugar nucleotide survey, together with a review of the literature, was used to define the routes to these important metabolites and to annotate relevant genes in the trypanosomatid genomes.

* Corresponding author. Mailing address: Division of Biological Chemistry and Drug Discovery, Wellcome Trust Biocentre, College of Life Sciences, University of Dundee, Dow St., Dundee DD1 5EH, Scotland, United Kingdom. Phone: 44-1382-384219. Fax: 44-1382-348896. E-mail: m.a.j.ferguson{at}dundee.ac.uk

{triangledown} Published ahead of print on 8 June 2007.

{dagger} Supplemental material for this article may be found at http://ec.asm.org/.

Eukaryotic Cell, August 2007, p. 1450-1463, Vol. 6, No. 8
1535-9778/07/$08.00+0     doi:10.1128/EC.00175-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



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