Developmental regulation of proline transport in Leishmania donovani.
Leishmania donovani are the causative agents of kala azar in humans. These organisms cycle between the proline-rich environment of the sand fly vector (extracellular promastigotes) and the sugar-rich condition in the mammalian host (intracellular amastigotes). Parasites have adapted to these extreme changes in proline concentrations: promastigotes utilize proline as a carbon source, whereas amastigotes utilize sugars and fatty acids. Previous studies have suggested that promastigotes and amastigotes express distinct proline transporters. However, the information available on these transporters is limited. In this work, proline transport was investigated in axenic L. donovani cultures. Three transport systems were identified: cation-dependent and -independent proline transporters in promastigotes (systems A and B, respectively) and a single cation-independent transporter in amastigotes (system C). Systems A and C have broad specificity to almost all amino acids and obtain optimum activity at acidic pH ranges (pH 6 and 5, respectively). System B is more specific to proline, as it is inhibited by only five amino acids. Temperature response analyses indicated that the transporters of both promastigotes and amastigotes perform best at 37 degrees C. The activity of system A during parasite differentiation was assessed. The transport activity of system A disappeared 3 days after promastigotes were induced to differentiate into amastigotes. In these cells, elevated temperature and acidic pH each suppressed the activity of system A. When amastigotes were induced to differentiate back into promastigotes, system A resumed its activity 24 h after differentiation was initiated. In conclusion, L. donovani obtain proline transport systems that are stage specific, regulated by both pH and temperature. This paper constitutes the first investigation of amino acid transport in axenic L. donovani.
Mazareb
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Department of Biology, Technion-Israel, Institute of Technology, Haifa, 32000, Israel.
Fu
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Zilberstein
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D026921:Amino Acid Transport Systems, Neutral; D000596:Amino Acids / Q000378:metabolism; D000818:Animals; D001692:Biological Transport / Q000187:drug effects; D006863:Hydrogen-Ion Concentration; D007893:Leishmania donovani / Q000187:drug effects / Q000254:growth & development / Q000378:metabolism*; D026901:Membrane Transport Proteins / Q000378:metabolism*; D011188:Potassium / Q000494:pharmacology; D011392:Proline / Q000378:metabolism*; D012964:Sodium / Q000494:pharmacology; D013379:Substrate Specificity
D026921:Amino Acid Transport Systems, Neutral; D000596:Amino Acids; D026901:Membrane Transport Proteins; C033082:proline transporter; D011392:Proline; D012964:Sodium; D011188:Potassium