Cloning of a lymphatic peptide/histidine transporter.
Although peptide transport across the plasma membrane has been characterized well in the kidney and the intestine, the functional relevance of this transport in other organs has not been addressed. Here we report the cloning of a cDNA for a novel peptide/histidine transporter found in the rat (rPHT2), whose mRNA is expressed mainly in the lymphatic system. rPHT2 encodes a protein of 582 amino acids and showed 49% identity with the brain PHT (PHT1) [Yamashita, Shimada, Guo, Sato, Kohmura, Hayakawa, Takagi and Tohyama (1997) J. Biol. Chem. 272, 10205-10211]. rPHT2 mRNA was abundant in lung, spleen and thymus, and detected faintly in brain, liver, adrenal gland and heart by Northern-blot analysis and reverse transcriptase PCR (RT-PCR). Intense signals for the gene were found in immunocytes using in situ hybridization. Ectopic expression of rPHT2 protein in HEK-293T cells and BHK cells was not found on the cell surface, but was found on the lysosomal membrane using light- and electron-microscopic analysis. Recombinant rPHT2 protein reconstituted into liposomes showed proton-dependent transport activity with histidine and histidyl-leucine. These findings suggest that rPHT2 is involved in the protein catabolic pathway in the lymphatic system.
Sakata
K
K
Department of Anatomy and Neuroscience, Graduate School of Medicine Osaka University, 2-2 Yamadaoka, Suita, Osaka 565-0871, Japan. sakata@anat2.med.osaka-u.ac.jp
Yamashita
T
T
Maeda
M
M
Moriyama
Y
Y
Shimada
S
S
Tohyama
M
M
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001692:Biological Transport; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism*; D002451:Cell Compartmentation; D018076:DNA, Complementary; D004151:Dipeptides / Q000378:metabolism*; D006639:Histidine / Q000378:metabolism*; D008081:Liposomes; D008208:Lymphatic System / Q000378:metabolism*; D008247:Lysosomes; D026901:Membrane Transport Proteins; D008969:Molecular Sequence Data; D051381:Rats; D017422:Sequence Analysis, DNA; D014018:Tissue Distribution
D002352:Carrier Proteins; D018076:DNA, Complementary; D004151:Dipeptides; D008081:Liposomes; D026901:Membrane Transport Proteins; C429144:Slc15a3 protein, rat; D006639:Histidine; C020357:histidylleucine