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Fatty Acid Uptake in African Trypanosomes
Author(s) -
Paul Kimberly Sue,
Vigueira Patrick,
Winston Brette,
McCall Barbara,
Cain Sarah,
Ligon Marianne,
McCallister Jennifer
Publication year - 2010
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.24.1_supplement.850.4
Subject(s) - trypanosoma brucei , bodipy , fatty acid , biochemistry , proteolysis , chemistry , fatty acid binding protein , glycolysis , biology , metabolism , enzyme , fluorescence , gene , physics , quantum mechanics
Trypanosoma brucei is a parasite that causes lethal disease African Sleeping Sickness in humans. T. brucei has two options to supply itself with fatty acids: synthesis via an ER‐localized elongase pathway, or uptake from the host. We used fluorescence‐ and radioactive‐based assays to characterize fatty acid uptake in T. brucei. Uptake of fluorescent BODIPY‐C12 fatty acid was concentration‐ and time‐dependent and appeared to be saturable. Uptake of BODIPY‐C5 was inefficient compared to BODIPY‐C12 and ‐C16, suggesting chain‐length specificity. Surface proteolysis with trypsin reduced BODIPY‐C12 uptake in a concentration‐dependent manner, suggesting a surface protein may be involved. Treatment with 2‐deoxyglucose or RNA interference of hexokinase reduced BODIPY‐C12 uptake, suggesting partial dependence on glycolysis (and perhaps ATP). Uptake of [3H]fatty acids complexed with BSA exhibited 10‐fold less uptake compared to uncomplexed fatty acids. Finally, RNA interference targeting a tandem array of 4 acyl‐CoA synthetases resulted in reduced BODIPY‐C12 and [3H]fatty acid uptake and subsequent cell death, suggesting at least one ACS gene is involved in uptake. Taken together, our data indicate that T. brucei use multiple pathways to meditate their robust fatty acid uptake. Elucidation of these mechanisms will enhance our understanding of the host‐parasite relationship and may lead to novel therapies.