An acid-activated nucleobase transporter from Leishmania major

Diana Ortiz, Marco Sanchez, Hans P. Koch, H. Peter Larson, Scott Landfear

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

Parasitic protozoa are unable to synthesize purines de novo and must import preformed purine nucleobases or nucleosides from their hosts. Leishmania major expresses two purine nucleobase transporters, LmaNT3 and LmaNT4. Previous studies revealed that at neutral pH, LmaNT3 is a broad specificity, high affinity nucleobase transporter, whereas LmaNT4 mediates the uptake of only adenine. Because LmaNT4 is required for optimal viability of the amastigote stage of the parasite that lives within acidified phagolysomal vesicles of mammalian macrophages, the function of this permease was examined under acidic pH conditions. At acidic pH, LmaNT4 acquires the ability to transport adenine, hypoxanthine, guanine, and xanthine with Km values in the micromolar range, indicating that this transporter is activated at low pH. Thus, LmaNT4 is an acid-activated purine nucleobase transporter that functions optimally under the physiological conditions the parasite is exposed to in the macrophage phagolysosome. In contrast, LmaNT3 functions optimally at neutral pH. Two-electrode voltage clamp experiments performed on LmaNT3 and LmaNT4 expressed in Xenopus oocytes revealed substrate-induced inward directed currents at acidic pH, and application of substrates induced acidification of the oocyte cytosol. These observations imply that LmaNT3 and LmaNT4 are nucleobase/proton symporters.

Original languageEnglish (US)
Pages (from-to)16164-16169
Number of pages6
JournalJournal of Biological Chemistry
Volume284
Issue number24
DOIs
StatePublished - Jun 12 2009

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Leishmania major
Macrophages
Adenine
Acids
Protozoa
Symporters
Purines
Hypoxanthine
Xanthine
Membrane Transport Proteins
Acidification
Clamping devices
Guanine
Substrates
Nucleosides
Oocytes
Protons
Parasites
Phagosomes
Electrodes

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology

Cite this

An acid-activated nucleobase transporter from Leishmania major. / Ortiz, Diana; Sanchez, Marco; Koch, Hans P.; Larson, H. Peter; Landfear, Scott.

In: Journal of Biological Chemistry, Vol. 284, No. 24, 12.06.2009, p. 16164-16169.

Research output: Contribution to journalArticle

Ortiz, Diana ; Sanchez, Marco ; Koch, Hans P. ; Larson, H. Peter ; Landfear, Scott. / An acid-activated nucleobase transporter from Leishmania major. In: Journal of Biological Chemistry. 2009 ; Vol. 284, No. 24. pp. 16164-16169.
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