Establishment and Characterization of Mammalian Cell Lines Stably Expressing Human L-Type Amino Acid Transporters

Journal of Pharmacological Sciences

Do Kyung Kim¹  Arthit Chairoungdua¹  Michael F. Wempe²  Naohiko Anzai¹  Hitoshi Endou¹  Yoshikatsu Kanai¹  Emiko Morimoto¹  Hye Won Choi¹ 

[1] Department of Pharmacology and Toxicology, Kyorin University School of Medicine, Japan;Department of Pharmacology, James H. Quillen College of Medicine, East Tennessee State University, USA

关键词: amino acid transporter;    system L (SL);    SLC7;    L-type amino acid transporter (LAT) 1;    LAT2;   

DOI  :  10.1254/jphs.08232FP

学科分类：药学

来源: Nihon Yakuri Gakkai Henshuubu / Japanese Pharmacological Society

【 摘 要 】

References(46)Cited-By(27)System L (SL), a basolateral amino acid transporter, transports large neutral amino acids (LNAAs) in a Na+-independent manner. Previously, we identified two isoforms of transporters: L-type amino acid transporter 1 (LAT1) and 2 (LAT2) and revealed their distinct substrate selectivity and transport properties. In this study, to establish more stable human LAT1 (hLAT1) and LAT2 (hLAT2) in vitro assay systems, we established mouse cell lines stably expressing hLAT1 (S2-LAT1) and hLAT2 (S2-LAT2). Real-time quantitative RT-PCR analysis revealed that S2-LAT1 and S2-LAT2 cells express hLAT1 and hLAT2 mRNAs at 20 – 1000-fold higher levels than those of endogenous mouse Lat1 and Lat2. S2-LAT1 and S2-LAT2 mediated [14C]L-leucine transport properties were measured and corresponded to results observed via Xenopus oocytes. Using these cells, the data demonstrate that hLAT1 and hLAT2 exhibit different characters in the acceptance of α-methyl amino acids and amino acid–related compounds with bulky side chains such as thyroid hormones and melphalan. S2-LAT1 and S2-LAT2 cells are expected to facilitate hLAT1 and hLAT2 substrate recognition research and contribute to drug development by providing an efficient assay system to screen for chemical compounds that interact with hLAT1 and hLAT2.

【 授权许可】

Unknown