Genetic validation of aldolase and glyceraldehyde-3-phosphate dehydrogenase as drug targets in Trypanosoma brucei

• Published in: Molecular and biochemical parasitology Vol. 169; no. 1; pp. 50 - 54
• Main Authors: Cáceres, Ana Judith, Michels, Paul A.M., Hannaert, Véronique
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.01.2010; Amsterdam: Elsevier
• Subjects: Aldolase; Animals; Cell Line; Fructose-Bisphosphate Aldolase - antagonists & inhibitors; Fructose-Bisphosphate Aldolase - genetics; Fructose-Bisphosphate Aldolase - metabolism; Gene Knockdown Techniques; Genetic Variation; Glucose - metabolism; Glyceraldehyde-3-phosphate dehydrogenase; Glyceraldehyde-3-Phosphate Dehydrogenases - genetics; Glyceraldehyde-3-Phosphate Dehydrogenases - metabolism; Glycolysis; Humans; Protozoan Proteins - genetics; Protozoan Proteins - metabolism; RNA Interference; RNA, Double-Stranded - genetics; RNA, Double-Stranded - therapeutic use; Trypanosoma brucei; Trypanosoma brucei brucei - drug effects; Trypanosoma brucei brucei - enzymology; Trypanosoma brucei brucei - genetics; Trypanosoma brucei brucei - growth & development; Trypanosomes; Trypanosomiasis, African - drug therapy; Trypanosomiasis, African - metabolism; Trypanosomiasis, African - parasitology; RNAi; PYK; RNA interference; PGAM; GPO; Trypanosomes; GPDH; PFK; PGK; IC50; Glycolysis; ALD; Glyceraldehyde-3-phosphate dehydrogenase; HXK; GAPDH; Aldolase
• ISSN: 0166-6851; 1872-9428
• DOI: 10.1016/j.molbiopara.2009.09.001

Aldolase (ALD) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) of Trypanosoma brucei are considered to be promising targets for chemotherapeutic treatment of African sleeping sickness, because glycolysis is the single source of ATP for the parasite when living in the human bloodstream. Moreover, these enzymes appeared to possess distinct kinetic and structural properties that have already been exploited for the discovery of effective and selective inhibitors with trypanocidal activity. Here we present an experimental, quantitative assessment of the importance of these enzymes for the glycolytic pathway. This was achieved by decreasing the concentrations of ALD and GAPDH by RNA interference. The effects of these knockdowns on parasite growth, levels of various enzymes and transcripts, enzyme activities and glucose consumption were studied. A partial depletion of ALD and GAPDH was already sufficient to rapidly kill the trypanosomes. An effect was also observed on the activity of some other glycolytic enzymes.