Molecular cloning and functional expression of a chicken intestinal peptide transporter (cPepT1) in Xenopus oocytes and Chinese hamster ovary cells.
To study peptide absorption in chickens, an intestinal peptide transporter cDNA (cPepT1) was isolated from a chicken duodenal cDNA library. The cDNA was 2914 bp long and encoded a protein of 714 amino acid residues with an estimated molecular size of 79.3 kDa and an isoelectric point of 7.48. cPepT1 protein is similar60% identical to PepT1 from rabbits, humans, mice, rats and sheep. Sixteen dipeptides, three tripeptides and four tetrapeptides that contained the essential amino acids Met, Lys and(or) Trp were used for functional analysis of cPepT1 in Xenopus oocytes and Chinese hamster ovary cells. For most di- and tripeptides tested, the substrate affinities were in the micromolar range, indicating that cPepT1 has high affinity for these peptides. Lys-Lys and Lys-Trp-Lys were exceptions, with substrate affinities in the millimolar range. Neither free amino acids nor tetrapeptides were transported by cPepT1. Northern blot analysis using a full-length cPepT1 cDNA as the probe demonstrated that cPepT1 is expressed strongly in the duodenum, jejunum and ileum, and at lower levels in kidney and ceca. The present study demonstrated for the first time the presence and functional characteristics of a peptide transport system from an avian species.
Chen
Hong
H
Department of Animal and Poultry Sciences, Virginia Polytechnic Institute and State University, Blacksburg, VA 24061, USA.
Pan
YuanXiang
Y
Wong
Eric A
EA
Bloomquist
Jeffrey R
JR
Webb
Kenneth E
KE
D000595:Amino Acid Sequence; D000818:Animals; D015152:Blotting, Northern; D016466:CHO Cells; D002352:Carrier Proteins / Q000737:chemistry / Q000235:genetics*; D002645:Chickens / Q000378:metabolism*; D003001:Cloning, Molecular; D006224:Cricetinae; D004151:Dipeptides / Q000378:metabolism; D004386:Duodenum / Q000737:chemistry; D005260:Female; D015870:Gene Expression; D007082:Ileum / Q000737:chemistry; D007526:Isoelectric Point; D007583:Jejunum / Q000737:chemistry; D007700:Kinetics; D026901:Membrane Transport Proteins; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009842:Oligopeptides / Q000378:metabolism; D009865:Oocytes; D012333:RNA, Messenger / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016415:Sequence Alignment; D017421:Sequence Analysis; D014018:Tissue Distribution; D014162:Transfection; D014982:Xenopus laevis
D002352:Carrier Proteins; D004151:Dipeptides; D026901:Membrane Transport Proteins; D009842:Oligopeptides; D012333:RNA, Messenger; C067802:intestinal peptide-proton cotransporter