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

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Author(s)

Abstract

System L (SL), a basolateral amino acid transporter, transports large neutral amino acids (LNAAs) in a Na<sup>+</sup>-independent manner. Previously, we identified two isoforms of transporters: <sc><font size = "-2">L</font></sc>-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 [<sup>14</sup>C]<sc><font size = "-2">L</font></sc>-leucine transport properties were measured and corresponded to results observed via <i>Xenopus</i> 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.<br>

Journal

  • Journal of Pharmacological Sciences

    Journal of Pharmacological Sciences 108(4), 505-516, 2008-12-20

    The Japanese Pharmacological Society

References:  46

Cited by:  3

Codes

  • NII Article ID (NAID)
    10025732972
  • NII NACSIS-CAT ID (NCID)
    AA11806667
  • Text Lang
    ENG
  • Article Type
    Journal Article
  • ISSN
    13478613
  • NDL Article ID
    9751894
  • NDL Source Classification
    ZS51(科学技術--薬学)
  • NDL Call No.
    Z53-D199
  • Data Source
    CJP  CJPref  NDL  J-STAGE 
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