2001 Fiscal Year Final Research Report Summary
Molecular mechanisms of drug transport across cell membrane
Project/Area Number |
11694310
|
Research Category |
Grant-in-Aid for Scientific Research (B)
|
Allocation Type | Single-year Grants |
Section | 一般 |
Research Field |
応用薬理学・医療系薬学
|
Research Institution | Kyorin University |
Principal Investigator |
ENDOU Hitoshi Kyorin Univ. School of Medicine, Dept. of Pharmacology & Toxicology, Professor, 医学部, 教授 (20101115)
|
Co-Investigator(Kenkyū-buntansha) |
HOSOYAMADA Makoto Kyorin Univ. School of Medicine, Dept. of Pharmacology & Toxicology, Lecturer, 医学部, 講師 (00291659)
TAKEDA Michio Kyorin Univ. School of Medicine, Dept. of Pharmacology & Toxicology, Associate Professor, 医学部, 助教授 (40255401)
KANAI Yoshikatsu Kyorin Univ. School of Medicine, Dept. of Pharmacology & Toxicology, Professor, 医学部, 教授 (60204533)
INATOMI Jun Kyorin Univ. School of Medicine, Dept. of Pharmacology & Toxicology, Research Associate, 医学部, 助手 (00311960)
KIM Do Kyung Kyorin Univ. School of Medicine, Dept. of Pharmacology & Toxicology, Research Associate, 医学部, 助手 (40327474)
|
Project Period (FY) |
1999 – 2001
|
Keywords | Drug transporter / Organic anion transporter / LAT family / Associating protein / Heterocllimer / Aromatic amino acid / Triiodethyronie / L-Dopa |
Research Abstract |
One of the remarkable features of the transporters responsible for the transmembrane transport of organic substances is their multispecificity in the substrate recognition, whose molecular mechanisms remain to be clarified. Even after the three-dimensional structures of the transporter proteins are solved by crystallography, it would be still difficult to understand the structural basis for the multispecific nature in the substrate recognition. Therefore, in the present investigation, we have performed structure-activity analysis using molecular pharmacological analysis. Neutral amino acid transporter LAT1 (L-type amino acid transporter 1) transports not only naturally-occurring L-amino acids nut also amino acid related drugs. In order to generate stably transected cell lines, we transected human LAT1 cDNA to S2 cells derived from mouse renal proximal tubule S2 segment of transgenic mice overexpressing SV40 large T antigen. We, then, examined the inhibitory effects of the compounds on the uptake of radiolabeled phenylalanine by the human LAT1-stable transfected cell line. We demonstrated that the phenylalanine uptake was inhibited by aromatic amino acid-related compounds such as L-dopa, alpha-methyldopa, triiodethyronine, thyroxine in a competitive manner. On the other hands phenylalanine-methylester, N-methylphenylalanine and dopamine had no effects on the LAT1-mediated transport. Computational analysis results indicated that presence of aromatic hydrophobic side chains as well as free carboxyl and free amino groups is essential to interacted with substrate binding site of LAT1. It was proved that the combination of the molecular pharmacological analysis and the cpmput ational analysis is the excellent means to reveal the structural basis of the substrate recognition of transporters.
|
Research Products
(14 results)
-
[Publications] Cha, S.H., Sekine, T., Fukushima, J., Kanai, Y., Kobayashi, Y., Goya, T, Endou, H: "Identification and characterization of human organic anion transporter 3 expressing predominanatly in the kidney"Mol.Pharmacol.. 59. 1277-1286 (2001)
Description
「研究成果報告書概要(和文)」より
-
[Publications] Kim, DK, Kanai, Y, Chairoungdua, A, Matsuo, H, Cha, SH, Endou, H: "Expression cloning of a Na^+-independent aromatic amino acid transporter with structural similarity to H^+/monocarboxylate transporters"J Biol.Chem.. 276(20). 17221-17228 (2001)
Description
「研究成果報告書概要(和文)」より
-
[Publications] Kim, J.Y., Kanai, Y, Chairoungdua, A., Cha, S.H., Matsuo, H., Kim, D.K., Inatomi, J., Sawa, H., Ida, Y., Endou, H: "Human cystine/glutamate transporter : cDNA cloning and upregulation by oxidative stress in glioma cells"Biochem.Biopys.Acta.. 1512. 335-344 (2001)
Description
「研究成果報告書概要(和文)」より
-
-
[Publications] Uchino, H., Kanai, Y., Kim D.K., Wenpe, MF., Chairoungdua, A., Moriinoto, E., Anders.M.W., Endou, H: "Transport of amino acid-related compounds mediated by L-type amino acid transorter 1 (LAT1): Insights into the mechanisms of substrate recognition"Mol.Pharmacol. 61. 729-737 (2001)
Description
「研究成果報告書概要(和文)」より
-
[Publications] Enomoto, A., Kimura, H., Chairoungdua, A., Shigeta, Y., Jutabha, P., Cha, S.H., Hosoyamada, M., Takeda, M., Sekine, T., Igarashi, T., Matsuo, H., Kikuchi, Y., Oda, T., Ichida, K., Hosoya, T., Shimokata, K., Niwa, T., Kanai, Y., Eendou, H: "Molecular identification of a renal urate/anion exchanger that regulates blood urate levels"Nature. (In press).
Description
「研究成果報告書概要(和文)」より
-
-
[Publications] Cha, S.H., Sekine, T., Fukushima, J., Kanai, Y., Kobayashi, Y., Goya, T., and Endou, H.: "Identification and characterization of human organic anion transporter 3 expressing predominantly in the kidney"Mol. Pharmacol. 59. 1277-1286 (2001)
Description
「研究成果報告書概要(欧文)」より
-
[Publications] Kim, DK, Kanai, Y, Chairoungdua, A, Matsuo, H, Cha, SH and Endou, H: "Expression cloning of a Na^+-independent aromatic amino acid transporter with structural similarity to H^+/monocarboxylate transporters"J. Biol. Chem.. 276(20). 17221-17228 (2001)
Description
「研究成果報告書概要(欧文)」より
-
[Publications] Kim, J.Y., Kanai, Y., Chairoungdua, A., Cha, S.H., Matsuo, H., Kim, D.K., Inatomi, J., Sawa, H., Ida, Y. and Endou, H.: "Human cystine/glutamate transporter: cDNA cloning and upregulation by oxidative stress in glioma cells"Biochem. Biopys. Acta. 1512. 335-344 (2001)
Description
「研究成果報告書概要(欧文)」より
-
-
[Publications] Uchino, H., Kanai, Y., Kim D.K., Wenpe, M.F., Chairoungdua, A., Morimoto, E., Anders, M.W. and Endou, H.: "Transport of amino acid-related compounds mediated by L-type amino acid transorter 1 (LAT1): Insights into the mechanisms of substrate recognition"Mol. Pharmacol. 61. 729-737 (2002)
Description
「研究成果報告書概要(欧文)」より
-
[Publications] Enomoto, A., Kimura, H., Chairoungdua, A., Shigeta, Y., Jutabha, P., Cha, S.H., Hosoyamada, M., Takeda, M., Sekine, T., Igarashi, T., Matsuo, H., Kikuchi, Y., Oda, T., Ichida, K., Hosoya, T., Shimokata, K., Niwa, T., Kanai Y. and Endou, H.: "Molecular identification of a renal urate-anion exchanger that regulates blood"urate levels. Nature 10 April 2002 (DOI 10. 1038/nature 742).
Description
「研究成果報告書概要(欧文)」より
-