Electrophysiological characteristics of the proton-coupled peptide transporter PEPT2 cloned from rat brain.
We have cloned a peptide transporter from rat brain and found it to be identical to rat kidney PEPT2. In the present study we characterize the transport function of the rat brain PEPT2, with special emphasis on electrophysiological properties and interaction with N-acetyl-L-aspartyl-L-glutamate (NAAG). When heterologously expressed in HeLa cells and in SK-N-SH cells, PEPT2 transports several dipeptides but not free amino acids in the presence of a proton gradient. NAAG competes with other peptides for the PEPT2-mediated transport process. When PEPT2 is expressed in Xenopus laevis oocytes, substrate-induced inward currents are detectable with dipeptides of differing charge in the presence of a proton gradient. Proton activation kinetics are similar for differently charged peptides. NAAG is a transportable substrate for PEPT2, as evidenced by NAAG-induced currents. The Hill coefficient for protons for the activation of the transport of differently charged peptides, including NAAG, is 1. Although the peptide-to-proton stoichiometry for negatively charged peptides is 1, the transport nonetheless is associated with transfer of positive charge into the oocyte, as indicated by peptide-induced inward currents.
Wang
H
H
Department of Biochemistry and Molecular Biology, Medical College of Georgia, Augusta, Georgia 30912-2100, USA.
Fei
Y J
YJ
Ganapathy
V
V
Leibach
F H
FH
D000596:Amino Acids / Q000378:metabolism*; D000818:Animals; D001692:Biological Transport; D001921:Brain / Q000502:physiology*; D002352:Carrier Proteins / Q000187:drug effects / Q000502:physiology*; D002460:Cell Line; D003001:Cloning, Molecular; D004151:Dipeptides / Q000378:metabolism* / Q000494:pharmacology; D006367:HeLa Cells; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007668:Kidney / Q000502:physiology; D007700:Kinetics; D018696:Neuroprotective Agents / Q000494:pharmacology; D011522:Protons; D051381:Rats; D011994:Recombinant Proteins / Q000187:drug effects / Q000378:metabolism; D027981:Symporters; D014162:Transfection
D000596:Amino Acids; D002352:Carrier Proteins; D004151:Dipeptides; D018696:Neuroprotective Agents; D011522:Protons; D011994:Recombinant Proteins; D027981:Symporters; C093694:hydrogen-coupled oligopeptide transporter PepT2; C027172:isospaglumic acid