PMID: 8956326Nov 1, 1996Paper

Human dipeptide transporter, hPEPT1, stably transfected into Chinese hamster ovary cells

Pharmaceutical Research
K M CovitzW Sadée

Abstract

A cDNA encoding the H(+)-coupled peptide transporter, hPEPT1, has previously been cloned from human ileum (8). The objective of this study was to establish a stably transfected cell line expressing hPEPT1 in mammalian cell culture. The hPEPT1 cDNA was subcloned into an expression vector carrying the CMV promoter and a neomycin resistance gene. This vector, pCDNA3-PEPT1, was transiently transfected into several cell lines to identify those capable of expressing PEPT1 transport function. CHO cells were selected and stably transfected with PEPT1 (CHO-PEPT1). Dipeptide transport activity was measured with 3H-Gly-Sar, in the presence and absence of inhibitors. The clonal cell line, CHO-PEPT1, displayed high transport activity. Dipeptide transport was sensitive to pH and specific for dipeptides and other small peptides. Peptidomimetic antibiotics, such as cephalexin, were competitors for peptide transport. The stably transfected cell line, CHO-PEPT1 exhibits enhanced transport over that of cell lines with native expression of PEPT1, and therefore, represents a useful tool for rapid screening of drugs that utilize the peptide transporter in the human intestine for absorption.

Citations

Jul 26, 2016·AMB Express·Andres Sánchez-KopperRalf Takors
May 17, 2003·The Journal of Pharmacology and Experimental Therapeutics·Christopher P LandowskiGordon L Amidon
Mar 22, 2003·The Journal of Pharmacology and Experimental Therapeutics·Hidekazu ToyobukuAkira Tsuji
Apr 23, 2005·Pharmaceutical Research·Sean EkinsPeter W Swaan
Nov 1, 2005·The Journal of Pharmacology and Experimental Therapeutics·Pascale AnderleWolfgang Sadée
May 23, 2002·The Journal of Pharmacology and Experimental Therapeutics·Hidekazu ToyobukuAkira Tsuji
Jan 20, 1998·Drug and Chemical Toxicology·M M Doherty, K S Pang
Mar 30, 2002·Advanced Drug Delivery Reviews·Eric Y ZhangPeter W Swaan
Oct 4, 2005·Protein Expression and Purification·Ryoichi SaitohMasayuki Tsuchiya
Mar 3, 2006·European Journal of Pharmaceutical Sciences : Official Journal of the European Federation for Pharmaceutical Sciences·Philipp SchlatterJuergen Drewe
Nov 14, 2006·Advanced Drug Delivery Reviews·Cheng ChangPeter W Swaan

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