Functional expression and characterization of a purine nucleobase transporter gene from Leishmania major

Marco Sanchez, Rob Tryon, Steven Pierce, Gayatri Vasudevan, Scott Landfear

Research output: Contribution to journalArticle

31 Citations (Scopus)

Abstract

Leishmania major, like all the other kinetoplastid protozoa, are unable to synthesize purines and rely on purine nucleobase and nucleoside acquisition across the parasite plasma membrane by specific permeases. Although, several genes have been cloned that encode nucleoside transporters in Leishmania and Trypanosoma brucei, much less progress has been made on nucleobase transporters, especially at the molecular level. The studies reported here have cloned and expressed the first gene for a L. major nucleobase transporter, designated LmaNT3. The LmaNT3 permease shows 33% identity to L. donovani nucleoside transporter 1.1 (LdNT1.1) and is, thus, a member of the equilibrative nucleoside transporter (ENT) family. ENT family members identified to date are nucleoside transporters, some of which also transport one or several nucleobases. Functional expression studies in Xenopus laevis oocytes revealed that LmaNT3 mediates high levels of uptake of hypoxanthine, xanthine, adenine and guanine. Moreover, LmaNT3 is an high affinity transporter with K m values for hypoxanthine, xanthine, adenine and guanine of 16.5±1.5, 8.5± 0.6, 8.5±1.1 and 8.8±4.0 μM, respectively. LmaNT3 is, thus, the first member of the ENT family identified in any organism that functions as a nucleobase rather than nucleoside or nucleoside/nucleobase transporter.

Original languageEnglish (US)
Pages (from-to)11-18
Number of pages8
JournalMolecular Membrane Biology
Volume21
Issue number1
DOIs
StatePublished - Jan 2004

Fingerprint

Nucleoside Transport Proteins
Leishmania major
Genes
Hypoxanthine
Xanthine
Membrane Transport Proteins
Guanine
Adenine
Purine Nucleosides
Purines
Trypanosoma brucei brucei
Leishmania
Xenopus laevis
purine
Nucleosides
Oocytes
Parasites
Cell Membrane

Keywords

  • Kinetoplastida
  • Leishmania
  • Nucleobase transporter
  • Purines

ASJC Scopus subject areas

  • Cell Biology

Cite this

Functional expression and characterization of a purine nucleobase transporter gene from Leishmania major. / Sanchez, Marco; Tryon, Rob; Pierce, Steven; Vasudevan, Gayatri; Landfear, Scott.

In: Molecular Membrane Biology, Vol. 21, No. 1, 01.2004, p. 11-18.

Research output: Contribution to journalArticle

Sanchez, Marco ; Tryon, Rob ; Pierce, Steven ; Vasudevan, Gayatri ; Landfear, Scott. / Functional expression and characterization of a purine nucleobase transporter gene from Leishmania major. In: Molecular Membrane Biology. 2004 ; Vol. 21, No. 1. pp. 11-18.
@article{57bf9acf67a444d88a9112e3d920bd04,
title = "Functional expression and characterization of a purine nucleobase transporter gene from Leishmania major",
abstract = "Leishmania major, like all the other kinetoplastid protozoa, are unable to synthesize purines and rely on purine nucleobase and nucleoside acquisition across the parasite plasma membrane by specific permeases. Although, several genes have been cloned that encode nucleoside transporters in Leishmania and Trypanosoma brucei, much less progress has been made on nucleobase transporters, especially at the molecular level. The studies reported here have cloned and expressed the first gene for a L. major nucleobase transporter, designated LmaNT3. The LmaNT3 permease shows 33{\%} identity to L. donovani nucleoside transporter 1.1 (LdNT1.1) and is, thus, a member of the equilibrative nucleoside transporter (ENT) family. ENT family members identified to date are nucleoside transporters, some of which also transport one or several nucleobases. Functional expression studies in Xenopus laevis oocytes revealed that LmaNT3 mediates high levels of uptake of hypoxanthine, xanthine, adenine and guanine. Moreover, LmaNT3 is an high affinity transporter with K m values for hypoxanthine, xanthine, adenine and guanine of 16.5±1.5, 8.5± 0.6, 8.5±1.1 and 8.8±4.0 μM, respectively. LmaNT3 is, thus, the first member of the ENT family identified in any organism that functions as a nucleobase rather than nucleoside or nucleoside/nucleobase transporter.",
keywords = "Kinetoplastida, Leishmania, Nucleobase transporter, Purines",
author = "Marco Sanchez and Rob Tryon and Steven Pierce and Gayatri Vasudevan and Scott Landfear",
year = "2004",
month = "1",
doi = "10.1080/0968768031000140845",
language = "English (US)",
volume = "21",
pages = "11--18",
journal = "Molecular Membrane Biology",
issn = "0968-7688",
publisher = "Informa Healthcare",
number = "1",

}

TY - JOUR

T1 - Functional expression and characterization of a purine nucleobase transporter gene from Leishmania major

AU - Sanchez, Marco

AU - Tryon, Rob

AU - Pierce, Steven

AU - Vasudevan, Gayatri

AU - Landfear, Scott

PY - 2004/1

Y1 - 2004/1

N2 - Leishmania major, like all the other kinetoplastid protozoa, are unable to synthesize purines and rely on purine nucleobase and nucleoside acquisition across the parasite plasma membrane by specific permeases. Although, several genes have been cloned that encode nucleoside transporters in Leishmania and Trypanosoma brucei, much less progress has been made on nucleobase transporters, especially at the molecular level. The studies reported here have cloned and expressed the first gene for a L. major nucleobase transporter, designated LmaNT3. The LmaNT3 permease shows 33% identity to L. donovani nucleoside transporter 1.1 (LdNT1.1) and is, thus, a member of the equilibrative nucleoside transporter (ENT) family. ENT family members identified to date are nucleoside transporters, some of which also transport one or several nucleobases. Functional expression studies in Xenopus laevis oocytes revealed that LmaNT3 mediates high levels of uptake of hypoxanthine, xanthine, adenine and guanine. Moreover, LmaNT3 is an high affinity transporter with K m values for hypoxanthine, xanthine, adenine and guanine of 16.5±1.5, 8.5± 0.6, 8.5±1.1 and 8.8±4.0 μM, respectively. LmaNT3 is, thus, the first member of the ENT family identified in any organism that functions as a nucleobase rather than nucleoside or nucleoside/nucleobase transporter.

AB - Leishmania major, like all the other kinetoplastid protozoa, are unable to synthesize purines and rely on purine nucleobase and nucleoside acquisition across the parasite plasma membrane by specific permeases. Although, several genes have been cloned that encode nucleoside transporters in Leishmania and Trypanosoma brucei, much less progress has been made on nucleobase transporters, especially at the molecular level. The studies reported here have cloned and expressed the first gene for a L. major nucleobase transporter, designated LmaNT3. The LmaNT3 permease shows 33% identity to L. donovani nucleoside transporter 1.1 (LdNT1.1) and is, thus, a member of the equilibrative nucleoside transporter (ENT) family. ENT family members identified to date are nucleoside transporters, some of which also transport one or several nucleobases. Functional expression studies in Xenopus laevis oocytes revealed that LmaNT3 mediates high levels of uptake of hypoxanthine, xanthine, adenine and guanine. Moreover, LmaNT3 is an high affinity transporter with K m values for hypoxanthine, xanthine, adenine and guanine of 16.5±1.5, 8.5± 0.6, 8.5±1.1 and 8.8±4.0 μM, respectively. LmaNT3 is, thus, the first member of the ENT family identified in any organism that functions as a nucleobase rather than nucleoside or nucleoside/nucleobase transporter.

KW - Kinetoplastida

KW - Leishmania

KW - Nucleobase transporter

KW - Purines

UR - http://www.scopus.com/inward/record.url?scp=1642542400&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=1642542400&partnerID=8YFLogxK

U2 - 10.1080/0968768031000140845

DO - 10.1080/0968768031000140845

M3 - Article

C2 - 14668134

AN - SCOPUS:1642542400

VL - 21

SP - 11

EP - 18

JO - Molecular Membrane Biology

JF - Molecular Membrane Biology

SN - 0968-7688

IS - 1

ER -