Molecular cloning and functional expression of a chicken intestinal peptide transporter (cPepT1) in Xenopus oocytes and Chinese hamster ovary cells

Hong Chen, Yuan Xiang Pan, Eric A. Wong, Jeffrey R. Bloomquist, Kenneth E. Webb

Research output: Contribution to journalArticle

Abstract

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 ∼60% 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.

Original languageEnglish (US)
Pages (from-to)387-393
Number of pages7
JournalJournal of Nutrition
Volume132
Issue number3
StatePublished - Mar 20 2002
Externally publishedYes

Fingerprint

Molecular Cloning
Xenopus
Cricetulus
Oocytes
Ovary
Chickens
Complementary DNA
Dipeptides
lysyl-tryptophyl-alpha-lysine
lysyllysine
Amino Acids
Peptides
Essential Amino Acids
Cecum
Isoelectric Point
Jejunum
Gene Library
Ileum
Duodenum
Northern Blotting

Keywords

  • Chickens
  • PepT1
  • Peptide
  • Transport

ASJC Scopus subject areas

  • Medicine (miscellaneous)
  • Nutrition and Dietetics

Cite this

Molecular cloning and functional expression of a chicken intestinal peptide transporter (cPepT1) in Xenopus oocytes and Chinese hamster ovary cells. / Chen, Hong; Pan, Yuan Xiang; Wong, Eric A.; Bloomquist, Jeffrey R.; Webb, Kenneth E.

In: Journal of Nutrition, Vol. 132, No. 3, 20.03.2002, p. 387-393.

Research output: Contribution to journalArticle

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