Transport of amino acids and GABA analogues via the human proton-coupled amino acid transporter, hPAT1: Characterization of conditions for affinity and transport experiments in Caco-2 cells.

Research output: Contribution to journal › Journal article › Research › peer-review

Standard

Transport of amino acids and GABA analogues via the human proton-coupled amino acid transporter, hPAT1 : Characterization of conditions for affinity and transport experiments in Caco-2 cells. / Larsen, Mie; Larsen, Birger Brodin; Frølund, Bente; Nielsen, Carsten Uhd.

In: European Journal of Pharmaceutical Sciences, Vol. 35, No. 1-2, 2008, p. 86-95.

Research output: Contribution to journal › Journal article › Research › peer-review

Harvard

Larsen, M, Larsen, BB, Frølund, B & Nielsen, CU 2008, 'Transport of amino acids and GABA analogues via the human proton-coupled amino acid transporter, hPAT1: Characterization of conditions for affinity and transport experiments in Caco-2 cells.', European Journal of Pharmaceutical Sciences, vol. 35, no. 1-2, pp. 86-95. https://doi.org/10.1016/j.ejps.2008.06.007

APA

Larsen, M., Larsen, B. B., Frølund, B., & Nielsen, C. U. (2008). Transport of amino acids and GABA analogues via the human proton-coupled amino acid transporter, hPAT1: Characterization of conditions for affinity and transport experiments in Caco-2 cells. European Journal of Pharmaceutical Sciences, 35(1-2), 86-95. https://doi.org/10.1016/j.ejps.2008.06.007

Vancouver

Larsen M, Larsen BB, Frølund B, Nielsen CU. Transport of amino acids and GABA analogues via the human proton-coupled amino acid transporter, hPAT1: Characterization of conditions for affinity and transport experiments in Caco-2 cells. European Journal of Pharmaceutical Sciences. 2008;35(1-2):86-95. https://doi.org/10.1016/j.ejps.2008.06.007

Author

Larsen, Mie ; Larsen, Birger Brodin ; Frølund, Bente ; Nielsen, Carsten Uhd. / Transport of amino acids and GABA analogues via the human proton-coupled amino acid transporter, hPAT1 : Characterization of conditions for affinity and transport experiments in Caco-2 cells. In: European Journal of Pharmaceutical Sciences. 2008 ; Vol. 35, No. 1-2. pp. 86-95.

Bibtex

@article{56207e90938011dd86a6000ea68e967b,
title = "Transport of amino acids and GABA analogues via the human proton-coupled amino acid transporter, hPAT1: Characterization of conditions for affinity and transport experiments in Caco-2 cells.",
abstract = "The objective of this study was to investigate transepithelial amino acid transport as a function of Caco-2 cell culture time. Furthermore, the objective was to investigate apical uptake characteristics of hPAT1-mediated transport under various experimental conditions. Apical amino acid uptake and transport studies were conducted in Caco-2 monolayers cultured for 4-28 days. Transepithelial transport of the prototypic hPAT1 (SLC36A1) substrates l-proline and glycine were maximal after 21-28 days in culture. Based on proton-dependency and substrate kinetics the major apical uptake and transport of Gly and Pro in Caco-2 cell monolayers is hPAT1-mediated. The apical uptake of Pro is decreased at apical hyperosmolarity conditions. Furthermore we identified the two GABA-analogues, muscimol and THPO as novel hPAT1 substrates. THPO had an affinity for hPAT1 of 11.3mM, whereas muscimol had one of the highest affinities for hPAT1 (1.7mM) reported. Our findings illustrate the suitability of the Caco-2 model for studying hPAT1-mediated transport. Furthermore, the affinity of THPO and muscimol underlines the possible importance of hPAT1 as a transporter for heterocyclic compounds consisting of a 3-isoxazolol moiety, which has been shown to function as a carboxylic acid bioisostere for substrates of the GABA receptor and transport systems.",
keywords = "Former Faculty of Pharmaceutical Sciences",
author = "Mie Larsen and Larsen, {Birger Brodin} and Bente Fr{\o}lund and Nielsen, {Carsten Uhd}",
note = "Keywords: hPAT1; Culture time; Amino acid transport; Osmolarity; GABA-analogues; Muscimol",
year = "2008",
doi = "10.1016/j.ejps.2008.06.007",
language = "English",
volume = "35",
pages = "86--95",
journal = "Norvegica Pharmaceutica Acta",
issn = "0928-0987",
publisher = "Elsevier",
number = "1-2",

}

RIS

TY - JOUR

T1 - Transport of amino acids and GABA analogues via the human proton-coupled amino acid transporter, hPAT1

T2 - Characterization of conditions for affinity and transport experiments in Caco-2 cells.

AU - Larsen, Mie

AU - Larsen, Birger Brodin

AU - Frølund, Bente

AU - Nielsen, Carsten Uhd

N1 - Keywords: hPAT1; Culture time; Amino acid transport; Osmolarity; GABA-analogues; Muscimol

PY - 2008

Y1 - 2008

N2 - The objective of this study was to investigate transepithelial amino acid transport as a function of Caco-2 cell culture time. Furthermore, the objective was to investigate apical uptake characteristics of hPAT1-mediated transport under various experimental conditions. Apical amino acid uptake and transport studies were conducted in Caco-2 monolayers cultured for 4-28 days. Transepithelial transport of the prototypic hPAT1 (SLC36A1) substrates l-proline and glycine were maximal after 21-28 days in culture. Based on proton-dependency and substrate kinetics the major apical uptake and transport of Gly and Pro in Caco-2 cell monolayers is hPAT1-mediated. The apical uptake of Pro is decreased at apical hyperosmolarity conditions. Furthermore we identified the two GABA-analogues, muscimol and THPO as novel hPAT1 substrates. THPO had an affinity for hPAT1 of 11.3mM, whereas muscimol had one of the highest affinities for hPAT1 (1.7mM) reported. Our findings illustrate the suitability of the Caco-2 model for studying hPAT1-mediated transport. Furthermore, the affinity of THPO and muscimol underlines the possible importance of hPAT1 as a transporter for heterocyclic compounds consisting of a 3-isoxazolol moiety, which has been shown to function as a carboxylic acid bioisostere for substrates of the GABA receptor and transport systems.

AB - The objective of this study was to investigate transepithelial amino acid transport as a function of Caco-2 cell culture time. Furthermore, the objective was to investigate apical uptake characteristics of hPAT1-mediated transport under various experimental conditions. Apical amino acid uptake and transport studies were conducted in Caco-2 monolayers cultured for 4-28 days. Transepithelial transport of the prototypic hPAT1 (SLC36A1) substrates l-proline and glycine were maximal after 21-28 days in culture. Based on proton-dependency and substrate kinetics the major apical uptake and transport of Gly and Pro in Caco-2 cell monolayers is hPAT1-mediated. The apical uptake of Pro is decreased at apical hyperosmolarity conditions. Furthermore we identified the two GABA-analogues, muscimol and THPO as novel hPAT1 substrates. THPO had an affinity for hPAT1 of 11.3mM, whereas muscimol had one of the highest affinities for hPAT1 (1.7mM) reported. Our findings illustrate the suitability of the Caco-2 model for studying hPAT1-mediated transport. Furthermore, the affinity of THPO and muscimol underlines the possible importance of hPAT1 as a transporter for heterocyclic compounds consisting of a 3-isoxazolol moiety, which has been shown to function as a carboxylic acid bioisostere for substrates of the GABA receptor and transport systems.

KW - Former Faculty of Pharmaceutical Sciences

U2 - 10.1016/j.ejps.2008.06.007

DO - 10.1016/j.ejps.2008.06.007

M3 - Journal article

C2 - 18621125

VL - 35

SP - 86

EP - 95

JO - Norvegica Pharmaceutica Acta

JF - Norvegica Pharmaceutica Acta

SN - 0928-0987

IS - 1-2

ER -

ID: 6445889