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Human intestinal H+/peptide cotransporter. Cloning, functional expression, and chromosomal localization

R Liang1, Y J Fei, P D Prasad

  • 1Department of Biochemistry and Molecular Biology, Medical College of Georgia, Augusta 30912-2100.

The Journal of Biological Chemistry
|March 24, 1995
PubMed
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Researchers identified a human gene encoding a peptide transporter crucial for absorbing dietary peptides in the small intestine. This transporter, located on chromosome 13, facilitates peptide uptake but not free amino acids.

Area of Science:

  • Molecular Biology
  • Human Physiology
  • Genetics

Background:

  • Mammalian small intestine utilizes a proton (H+)-coupled peptide transporter for absorbing small peptides from digested dietary proteins.
  • Previous isolation of a rabbit intestinal H+/peptide cotransporter cDNA provided a basis for further research.

Purpose of the Study:

  • To identify and characterize the human homolog of the H+/peptide cotransporter.
  • To investigate the functional properties and chromosomal localization of the cloned human transporter.

Main Methods:

  • Screening of a human intestinal cDNA library using a probe derived from the rabbit H+/peptide cotransporter cDNA.
  • Expression of the identified cDNA in HeLa cells and Xenopus laevis oocytes to assess peptide transport activity.
  • Bioinformatic analysis of the predicted protein sequence and chromosomal assignment studies (somatic cell hybrid analysis, in situ hybridization).

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Main Results:

  • A human cDNA clone was identified, which upon expression, induced H(+)-dependent peptide transport activity.
  • The encoded protein (708 amino acids) features 12 membrane-spanning domains and accepts dipeptides, tripeptides, and amino beta-lactam antibiotics, but not free amino acids.
  • High homology (81% identity, 92% similarity) was observed with the rabbit ortholog, but weak homology with bacterial and yeast peptide transporters.
  • The human H+/peptide cotransporter gene was localized to chromosome 13q33-->q34.

Conclusions:

  • A functional human H+/peptide cotransporter has been cloned and characterized, playing a key role in intestinal peptide absorption.
  • The human transporter shares significant structural and functional similarities with its rabbit counterpart but differs from microbial and yeast peptide transporters.
  • The gene's localization to chromosome 13 provides a genetic marker for further studies on peptide transport and related physiological processes.