Characterization of Two Mitochondrial Flavin Adenine Dinucleotide-Dependent Glycerol-3-Phosphate Dehydrogenases in Trypanosoma brucei

0421249 - BC 2014 RIV US eng J - Článek v odborném periodiku
Škodová, Ingrid - Verner, Zdeněk - Bringaud, F. - Fabian, P. - Lukeš, Julius - Horváth, A.
Characterization of Two Mitochondrial Flavin Adenine Dinucleotide-Dependent Glycerol-3-Phosphate Dehydrogenases in Trypanosoma brucei.
Eukaryotic Cell. Roč. 12, č. 12 (2013), s. 1664-1673. ISSN 1535-9778
Grant CEP: GA ČR(CZ) GAP305/11/2179; GA ČR GD206/09/H026; GA MŠMT LH12104
Institucionální podpora: RVO:60077344
Klíčová slova: alternative NADH dehydrogenase * inducible expression system * blood-stream forms * complex-I * procyclic trypanosomes * sleeping sickness * oxidase * localization * metabolism * cycle
Kód oboru RIV: EB - Genetika a molekulární biologie
Impakt faktor: 3.179, rok: 2013

Glycerol-3-phosphate dehydrogenases (G3PDHs) constitute a shuttle that serves for regeneration of NAD(+) reduced during glycolysis. This NAD-dependent enzyme is employed in glycolysis and produces glycerol-3-phosphate from dihydroxyacetone phosphate, while its flavin adenine dinucleotide (FAD)-dependent homologue catalyzes a reverse reaction coupled to the respiratory chain. Trypanosoma brucei possesses two FAD-dependent G3PDHs. While one of them (mitochondrial G3PDH [mtG3PDH]) has been attributed to the mitochondrion and seems to be directly involved in G3PDH shuttle reactions, the function of the other enzyme (putative G3PDH [putG3PDH]) remains unknown. In this work, we used RNA interference and protein overexpression and tagging to shed light on the relative contributions of both FAD-G3PDHs to overall cellular metabolism. Our results indicate that mtG3PDH is essential for the bloodstream stage of T. brucei, while in the procyclic stage the enzyme is dispensable. Expressed putG3PDH-V5 was localized to the mitochondrion, and the data obtained by digitonin permeabilization, Western blot analysis, and immunofluorescence indicate that putG3PDH is located within the mitochondrion.
Trvalý link: http://hdl.handle.net/11104/0227696