A novel H+-coupled oligopeptide transporter (OPT3) from Caenorhabditis elegans with a predominant function as a H+ channel and an exclusive expression in neurons

You Jun Fei, Michael F. Romero, Michael Krause, Jin Cai Liu, Wei Huang, Vadivel Ganapathy, Frederick H. Leibach

Research output: Contribution to journalArticle

25 Scopus citations

Abstract

We have cloned and functionally characterized a novel, neuron-specific, H+-coupled oligopeptide transporter (OPT3) from Caenorhabditis elegans that functions predominantly as a H+ channel. The opt3 gene is ~4.4 kilobases long and consists of 13 exons. The cDNA codes for a protein of 701 amino acids with 11 putative transmembrane domains. When expressed in mammalian cells and in Xenopus laevis oocytes, OPT3 cDNA induces H+-coupled transport of the dipeptide glycylsarcosine. Electrophysiological studies of the transport function of OPT3 in Xenopus oocytes show that this transporter, although capable of mediating H+-coupled peptide transport, functions predominantly as a H+ channel. The H+ channel activity of OPT3 is ~3-4- fold greater than the H+/peptide cotransport activity as determined by measurements of H+ gradient-induced inward currents in the absence and presence of the dipeptide using the two-microelectrode voltage clamp technique. A downhill influx of H+ was accompanied by a large intracellular acidification as evidenced from the changes in intracellular pH using an ion- selective microelectrode. The H+ channel activity exhibits a K0.5(H) of 1.0 μM at a membrane potential of-50 mV. At the level of primary structure, OPT3 has moderate homology with OPT1 and OPT2, two other H+-coupled oligopeptide transporters previously cloned from C. elegans. Expression studies using the opt3::gfp fusion constructs in transgenic C. elegans demonstrate that opt3 gene is exclusively expressed in neurons. OPT3 may play an important physiological role as a pH balancer in the maintenance of H+ homeostasis in C. elegans.

Original languageEnglish (US)
Pages (from-to)9563-9571
Number of pages9
JournalJournal of Biological Chemistry
Volume275
Issue number13
DOIs
StatePublished - Mar 31 2000

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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