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  • Differential kinetic activities of glycerol kinase among African trypanosome species: phylogenetic and therapeutic implications.

Differential kinetic activities of glycerol kinase among African trypanosome species: phylogenetic and therapeutic implications.

The Journal of veterinary medical science (2010-12-29)
Mitsuko Ohashi-Suzuki, Yoshisada Yabu, Shigeru Ohshima, Kosuke Nakamura, Yasutoshi Kido, Kimitoshi Sakamoto, Kiyoshi Kita, Nobuo Ohta, Takashi Suzuki
RESUMEN

African trypanosome species are causative agents for sleeping sickness in humans and nagana disease in cattle. Trypanosoma brucei can generate ATP via a reverse reaction with glycerol kinase (GK) when alternative oxidase (AOX) is inhibited; thus, GK is considered to be a crucial target for chemotherapy combined with AOX. However, the energy metabolism systems of African trypanosome species other than T. brucei are poorly understood. Thus, GK genes were surveyed from genome databases and cloned by PCR from T. vivax and T. congolense. Then, recombinant GK proteins (rGK) of T. vivax, T. congolense and T. brucei were expressed and purified. Kinetic analysis of these rGK proteins revealed that the K(m) values of T. congolense rGK for ADP and G-3-P substrates were lower than those of T. vivax and T. brucei. The expression level of GK molecules was highest in T. congolense cells and lowest in T. vivax cells. Based on these results, effective combination dosages of ascofuranone, a specific inhibitor of AOX, and glycerol, an inhibitor of the GK reverse reaction, were determined by using in vitro-cultured trypanosome cells.

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Sigma-Aldrich
Glycerokinase from Cellulomonas sp., lyophilized powder, 25-75 units/mg protein
Sigma-Aldrich
Glycerokinase from Escherichia coli, 300-600 units/mL, ammonium sulfate suspension
Sigma-Aldrich
Glycerokinase from Bacillus stearothermophilus, buffered aqueous solution, ≥75 units/mg protein (biuret)
Sigma-Aldrich
Glycerokinase from Escherichia coli, lyophilized powder, 40-100 units/mg protein