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

We have cloned and functionally characterized a novel, neuron-specific, H+- coupled oligopeptide transporter (OPT3) from Caenorhabditis elegans that fu nctions predominantly as a H+ channel. The opt3 gene is similar to 4.4 kilo bases long and consists of 13 exons. The cDNA codes for a protein of 701 am ino acids with 11 putative transmembrane domains. When expressed in mammali an cells and in Xenopus laevis oocytes, OPT3 cDNA induces H+-coupled transp ort of the dipeptide glycylsarcosine, Electrophysiological studies of the t ransport function of OPT3 in Xenopus oocytes show that this transporter, al though capable of mediating H+-coupled peptide transport, functions predomi nantly as a H+ channel. The H+ channel activity of OPT3 is similar to 3-4-f old greater than the H+/peptide cotransport activity as determined by measu rements of H+ gradient-induced inward currents in the absence and presence of the dipeptide using the two-microelectrode voltage clamp technique, A do wnhill influx of H+ was accompanied by a large intracellular acidification as evidenced from the changes in intracellular pH using an ion-selective mi croelectrode. The H+ channel activity exhibits a K-0.5(H) of 1.0 mu M at a membrane potential of -50 mV. At the level of primary structure, OPT3 has m oderate homology with OPT1 and OPT2, two other Hf-coupled oligopeptide tran sporters previously cloned from C. elegans, Expression studies using the op t3::gfp fusion constructs in transgenic C. elegans demonstrate that opt3 ge ne is exclusively expressed in neurons. OPT3 may play an important physiolo gical role as a pH balancer in the maintenance of H+ homeostasis in C. eleg ans.