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Leishmania LABCG2 transporter is involved in ATP-dependent transport of thiols

AuthorsPerea, A.; Manzano, J.I.; Kimura, Y.; Ueda, K.; Castanys, S.; Gamarro, F.
KeywordsABC transporters
oxidative stress
Issue Date2018
PublisherPortland Press
CitationBiochemical Journal 475: 87- 97 (2018)
AbstractThe Leishmania LABCG2 transporter has a key role in the redox metabolism of these protozoan parasites. Recently, the involvement of LABCG2 in virulence, autophagy and oxidative stress has been described. Null mutant parasites for LABCG2 present an increase in the intracellular levels of glutathione (GSH) and trypanothione [T(SH)]. On the other hand, parasites overexpressing LABCG2 transporter export non-protein thiols to the extracellular medium. To explore if LABCG2 may mediate an active transport of non-protein thiols, the effect of these molecules on ATPase activity of LABCG2 as well as the ability of LABCG2 to transport them was determined using a baculovirus-Sf9 insect cell system. Our results indicate that all thiols tested [GSH, T(SH)] as well as their oxidized forms GSSG and TS (trypanothione disulfide) stimulate LABCG2-ATPase basal activity. We have measured the transport of [H]-GSH in inside-out Sf9 cell membrane vesicles expressing LABCG2-GFP (green fluorescence protein), finding that LABCG2 was able to mediate a rapid and concentration-dependent uptake of [H]-GSH in the presence of ATP. Finally, we have analyzed the ability of different thiol species to compete for this uptake, T(SH) and TS being the best competitors. The IC value for [H]-GSH uptake in the presence of increasing concentrations of T(SH) was less than 100 μM, highlighting the affinity of this thiol for LABCG2. These results provide the first direct evidence that LABCG2 is an ABC transporter of reduced and oxidized non-protein thiols in Leishmania, suggesting that this transporter can play a role in the redox metabolism and related processes in this protozoan parasite.
Publisher version (URL)http://dx.doi.org/10.1042/BCJ20170685
Identifiersdoi: 10.1042/BCJ20170685
e-issn: 1470-8728
issn: 0264-6021
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