Primary structure genomic organization, and functional and electrogenic characteristics of human system N 1, a Na+ - and H+ -coupled glutamine transporter

You Jun Fei, Mitsuru Sugawara, Takeo Nakanishi, Wei Huang, Haiping Wang, Puttur D Prasad, Frederick H. Leibach, Vadivel Ganapathy

Research output: Contribution to journalArticle

89 Scopus citations

Abstract

We have cloned the human Na+- and H+-coupled amino acid transport system N (hSN1) from HepG2 liver cells and investigated its functional characteristics. Human SN1 protein consists of 504 amino acids and shows high homology to rat SN1 and rat brain glutamine transporter (GlnT). When expressed in mammalian cells, the transport function of human SN1 could be demonstrated with glutamine as the substrate in the presence of LiC1 (instead of NaC1) and cysteine. The transport activity was saturable, pH-sensitive, and specific for glutamine, histidine, asparagine, and alanine. Analysis of Li+ activation kinetics showed a Li+:glutamine stoichiometry of 2:1. When expressed in Xenopus laevis oocytes, the transport of glutamine or asparagine via human SN1 was associated with inward currents under voltage-clamped conditions. The transport function, monitored as glutamine- or asparagine-induced currents, was saturable, Na+-dependent, Li+-tolerant, and pH-sensitive. The transport cycle was associated with the involvement of more than one Na+ ion. Uptake of asparagine was directly demonstrable in these oocytes by using radiolabeled substrate, and this uptake was inhibited by merebrane depolarization. In addition, simultaneous measurement of asparagine influx and charge influx in the same oocyte yielded an asparagine:charge ratio of 1. These data suggest that SN1 mediates the influx of two Na+ and one amino acid substrate per transport cycle coupled to the efflux of one H+, rendering the transport process electrogenic.

Original languageEnglish (US)
Pages (from-to)23707-23717
Number of pages11
JournalJournal of Biological Chemistry
Volume275
Issue number31
DOIs
StatePublished - Aug 4 2000

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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