Overexpression of catalytic subunit p110 alpha of phosphatidylinositol 3-kinase increases glucose transport activity with translocation of glucose transporters in 3T3-L1 adipocytes

被引:156
作者
Katagiri, H
Asano, T
Ishihara, H
Inukai, K
Shibasaki, Y
Kikuchi, M
Yazaki, Y
Oka, Y
机构
[1] ASAHI LIFE FDN,INST ADULT DIS,SHINJUKU KU,TOKYO 160,JAPAN
[2] YAMAGUCHI UNIV,SCH MED,DEPT INTERNAL MED 3,UBE,YAMAGUCHI 755,JAPAN
关键词
D O I
10.1074/jbc.271.29.16987
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
To elucidate the mechanisms of phosphatidylinositol (PI) 3-kinase involvement in insulin-stimulated glucose transport activity, the epitope-tagged p110 alpha subunit of PI 3-kinase was overexpressed in 3T3-L1 adipocytes using an adenovirus-mediated gene transduction system, Overexpression of p110 alpha was confirmed by immunoblot using anti-tagged epitope antibody, p110 alpha overexpression induced a 2.5-fold increase in PI 3-kinase activity associated with its regulatory subunits in the basal state, an increase exceeding that of the maximally insulin-stimulated control cells, while PI 3-kinase activity associated with phosphotyrosyl protein was only modestly elevated, Overexpression of p110 alpha induced an approximately 14-fold increase in the basal glucose transport rate, which was also greater than that observed in the stimulated control. No apparent difference was observed in the cellular expression level of either GLUT1 or GLUT4 proteins between control and p110 alpha-overexpressing 3T3-L1 adipocytes, Subcellular fractionation revealed translocation of glucose transporters from intracellular to plasma membranes in basal p110 alpha-overexpressing cells, The translocation of GLUT4 protein to the plasma membrane was further confirmed using a membrane sheet assay, These findings indicate that an increment in PI 3-kinase activity induced by overexpression of p110 alpha of PI 3-kinase stimulates glucose transport activity with translocation of glucose transporters, i,e,, mimics the effect of insulin.
引用
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页码:16987 / 16990
页数:4
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