mRNA distribution and membrane localization of the OAT‐K1 organic anion transporter in rat renal tubules

FEBS Letters

Masuda, Satohiro¹  Inui, Ken-ichi¹  Tomita, Kimio²  Saito, Hideyuki¹  Nonoguchi, Hiroshi² 

[1] Department of Pharmacy, Kyoto University Hospital, Faculty of Medicine, Kyoto University, Sakyo-ku, Kyoto 606-01, Japan;Third Department of Internal Medicine, Kumamoto University School of Medicine, 1-1-1 Honjo, Kumamoto, Kumamoto 860, Japan

关键词: Organic anion transporter;    Methotrexate;    Microdissection;    Brush-border membrane;    Rat kidney;    oatp;    organic anion transporting polypeptide;    PCR;    polymerase chain reaction;    RT;    reverse transcription;    Tris;    2-amino-2-(hydroxymethyl)-1;    3-propanediol;    PMSF;    phenylmethylsulfonyl fluoride;    PVDF;    polyvinylidene difluoride;   

DOI  :  10.1016/S0014-5793(97)00314-1

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

OAT-K1, a renal organic anion transporter, which mediates methotrexate uptake, was analyzed for mRNA distribution along microdissected nephron segments and the immunolocalization in isolated plasma membranes from rat kidney. By using a reverse transcription-coupled PCR, OAT-K1 mRNA was detected predominantly in the superficial and juxtamedullary proximal straight tubules. Western blotting with antiserum for OAT-K1 revealed that the transporter protein with the apparent molecular mass of 40 kDa was expressed exclusively in the brush-border membranes from rat kidney. These findings suggest that the OAT-K1 is localized in the brush-border membranes of the renal proximal straight tubules.

【 授权许可】

Unknown