Targeting of GLUT1-GLUT5 chimeric proteins in the polarized cell line Caco-2

Kouichi Inukai, Kuniaki Takata, Tomoichiro Asano, Hideki Katagiri, Hisamitsu Ishihara, Mitsuhiro Nakazaki, Yasushi Fukushima, Yoshio Yazaki, Masatoshi Kikuchi, Yoshitomo Oka

研究成果: Article査読

13 被引用数 (Scopus)

抄録

Caco-2, a human differentiated intestinal epithelial cell line, is a promising model for investigating the mechanism of polarized targeting of apical and basolateral membrane proteins. We stably transfected rat GLUT5 cDNA and rabbit GLUT1 cDNA into Caco-2 cells with an expression vector. Immunohistochemical study revealed that the GLUT5 protein expressed was localized at apical membranes and that the GLUT1 expressed was present primarily in the basolateral membranes of cells grown on permeable support. Next, to investigate the domain responsible for determining apical vs. basolateral sorting in glucose transporters, we prepared several GLUT1-GLUT5 chimeric cDNAs and transfected them into Caco-2 cells. A GLUT1 [N terminus- sixth transmembrane domain (TM6)]-GLUT5 [intracellular loop (IL)~C terminus] chimera was observed exclusively at the apical membrane, while GLUT1 (N terminus~IL)-GLUT5 (TM7~C terminus) and GLUT1 (N terminus~TM12)-GLUT5 (C- terminal domain) chimeras were observed mainly at the basolateral membrane, a localization similar to that of GLUT1. Moreover, using a recombinant adenovirus expression system, we expressed a GLUT5 (N terminus~TM6) - GLUTI(IL)-GLUTS(TM7~C-terminus)chimera, which was observed at the basolateral membrane. Based on these results, the C-terminal domain does not determine isoform-specific targeting of GLUT1 and GLUT5. Rather, it is the intracellular loop in glucose transporters that appears to play a pivotal role in apical-basolateral sorting signals in Caco-2 cells.

本文言語English
ページ(範囲)442-449
ページ数8
ジャーナルMolecular Endocrinology
11
4
DOI
出版ステータスPublished - 1997
外部発表はい

ASJC Scopus subject areas

  • 分子生物学
  • 内分泌学

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