Functional characterization of the peptide transporter PEPT2 in primary cultures of human upper airway epithelium

Praveen M. Bahadduri, Vanessa M. D'Souza, Julia K. Pinsonneault, Wolfgang Sadée, Shenying Bao, Daren L. Knoell, Peter W. Swaan

Research output: Contribution to journalArticlepeer-review

30 Scopus citations

Abstract

This study characterizes the expression and function of the peptide transporter hPepT2 (SLC15A2) in differentiated primary cultures of human upper airway lung epithelia obtained from six human donors. Genotype analysis of a SNP in exon 15 of hPepT2 genotypes in six donors revealed an expected distribution of the two main variants present at similar frequency (two AA homozygotes, two BB homozygotes, and two AB heterozygotes). Real-time PCR analysis of the hPepT2 mRNA message revealed no significant differences among genotypes. hPEPT2 was expressed on the apical membrane in all donor specimens, demonstrated by cell surface biotinylation and Western analysis (104 kD). We then compared transepithelial transport of the prototypical substrate 3H- glycylsarcosine in all donor cultures in the absence and presence of known inhibitors of hPEPT2 to ascertain the phenotype of functionally expressed hPepT2 in the upper airway epithelium. An array of inhibitors included dipeptides, β-lactam antibiotics, bestatin, and ACE inhibitors. hPEPT2 exhibited saturable Michaelis-Menten-type kinetic parameters for GlySar, corroborating previously reported values for KT and Jmax. Donor-to-donor variation of transport for different substrates did not correlate with hPepT2 haplotypes in this sample cohort. These findings demonstrate functional hPEPT2 transporter expression in primary cultures of human lung epithelial cells. hPEP72-mediated transport could serve as a strategy for noninvasive systemic delivery of peptides and peptidomimetics drugs.

Original languageEnglish (US)
Pages (from-to)319-325
Number of pages7
JournalAmerican journal of respiratory cell and molecular biology
Volume32
Issue number4
DOIs
StatePublished - Apr 2005

Keywords

  • Drug transport
  • Genotype
  • Peptide transport
  • Polymorphism

ASJC Scopus subject areas

  • Molecular Biology
  • Pulmonary and Respiratory Medicine
  • Clinical Biochemistry
  • Cell Biology

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