| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
|
|
|
|||||||
|
|||||||||
The FASEB Journal, Vol 9, 1489-1496, Copyright © 1995 by The Federation of American Societies for Experimental Biology
RESEARCH COMMUNICATIONS |
M Brandsch, C Brandsch, PD Prasad, V Ganapathy, U Hopfer and FH Leibach
Department of Biochemistry and Molecular Biology, Medical College of Georgia, Augusta 30912, USA.
In this study we describe for the first time the identification of a renal cell line that expresses the kidney-specific high-affinity H+/peptide cotransport system. The kidney cell line SKPT-0193 C1.2 was obtained by SV40 transformation of rat proximal tubular cells. The transport of the dipeptide glycylsarcosine (Gly-Sar) was studied in this cell line grown as a confluent monolayer on impermeable plastic supports. Uptake of the dipeptide was rapid and was stimulated sixfold by an inwardly directed H+ gradient, with optimal uptake occurring at an extracellular pH of 6.0. The uptake was markedly reduced by the protonophore carbonyl cyanide 4-(trifluoromethoxy) phenylhydrazone whether measured at pH 7.5 or 6.0. Intracellular acidification of the cells by NH4Cl prepulse also reduced the uptake of glycylsarcosine. The dipeptide uptake was found to be mediated by a high-affinity transport system with a Michaelis-Menten constant (Kt) of 67 +/- 2 microM and a maximal transport velocity of 1.20 +/- 0.02 nmol.10 min-1.mg protein-1. Studied over a concentration range of 5 microM to 5 mM, there was no evidence for a second saturable transport component. Di- and tripeptides, but not glycine, were strong inhibitors of glycylsarcosine uptake, indicating that these peptides also interact with the transport system with high affinity. Northern blot analysis of poly(A)+RNA from these cells using cDNA probes specific for the human intestinal peptide transporter (PEPT 1) or the human kidney-specific peptide transporter (PEPT 2) revealed that the transport system expressed in these cells is PEPT 2. It is concluded that the SKPT-0193 C1.2 cell line constitutively expresses the kidney-specific high-affinity H+/peptide cotransporter described in the proximal tubular epithelial cells of the normal kidney.
This article has been cited by other articles:
|
|
 |
|
  A. Taranta, S. Petrini, A. Palma, L. Mannucci, M. J. Wilmer, V. De Luca, F. Diomedi-Camassei, S. Corallini, F. Bellomo, L. P. van den Heuvel, et al. Identification and subcellular localization of a new cystinosin isoform Am J Physiol Renal Physiol, May 1, 2008; 294(5): F1101 - F1108. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. M. Ocheltree, H. Shen, Y. Hu, R. F. Keep, and D. E. Smith Role and Relevance of Peptide Transporter 2 (PEPT2) in the Kidney and Choroid Plexus: In Vivo Studies with Glycylsarcosine in Wild-Type and PEPT2 Knockout Mice J. Pharmacol. Exp. Ther., October 1, 2005; 315(1): 240 - 247. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. A. Bravo, C. U. Nielsen, J. Amstrup, S. Frokjaer, and B. Brodin Epidermal growth factor decreases PEPT2 transport capacity and expression in the rat kidney proximal tubule cell line SKPT0193 cl.2 Am J Physiol Renal Physiol, February 1, 2004; 286(2): F385 - F393. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Neumann and M. Brandsch delta -Aminolevulinic Acid Transport in Cancer Cells of the Human Extrahepatic Biliary Duct J. Pharmacol. Exp. Ther., April 1, 2003; 305(1): 219 - 224. [Abstract] [Full Text]
|
|
|
|
 |
|
 I. Knutter, I. Rubio-Aliaga, M. Boll, G. Hause, H. Daniel, K. Neubert, and M. Brandsch H+-peptide cotransport in the human bile duct epithelium cell line SK-ChA-1 Am J Physiol Gastrointest Liver Physiol, July 1, 2002; 283(1): G222 - G229. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. Theis, I. Knutter, B. Hartrodt, M. Brandsch, G. Kottra, K. Neubert, and H. Daniel Synthesis and Characterization of High Affinity Inhibitors of the H+/Peptide Transporter PEPT2 J. Biol. Chem., February 22, 2002; 277(9): 7287 - 7292. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 C. Shu, H. Shen, U. Hopfer, and D. E. Smith Mechanism of Intestinal Absorption and Renal Reabsorption of an Orally Active Ace Inhibitor: Uptake and Transport of Fosinopril in Cell Cultures Drug Metab. Dispos., October 1, 2001; 29(10): 1307 - 1315. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
N. S. Teuscher, A. Novotny, R. F. Keep, and D. E. Smith Functional Evidence for Presence of PEPT2 in Rat Choroid Plexus: Studies with Glycylsarcosine J. Pharmacol. Exp. Ther., August 1, 2000; 294(2): 494 - 499. [Abstract] [Full Text]
|
|
|
|
|
|
H. Shen, D. E. Smith, T. Yang, Y. G. Huang, J. B. Schnermann, and F. C. Brosius III Localization of PEPT1 and PEPT2 proton-coupled oligopeptide transporter mRNA and protein in rat kidney Am J Physiol Renal Physiol, May 1, 1999; 276(5): F658 - F665. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 U. Wenzel, D. Diehl, M. Herget, and H. Daniel Endogenous expression of the renal high-affinity H+-peptide cotransporter in LLC-PK1 cells Am J Physiol Cell Physiol, December 1, 1998; 275(6): C1573 - C1579. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
W. Jin and U. Hopfer Dipeptide-induced Cl- secretion in proximal tubule cells Am J Physiol Cell Physiol, November 1, 1997; 273(5): C1623 - C1631. [Abstract] [Full Text] [PDF]
|
|
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
