Effects of wortmannin on glucose uptake and protein kinase C activity in rat adipocytes.
Wortmannin is known to be an inhibitor of myosin light chain kinase and phosphatidylinositol 3-kinase (PI 3-kinase) (J. Biol. Chem. 268, 25846, 1993). We studied the effects of wortmannin on insulin- and 12-O-tetradecanoylphorbol 13-acetate (TPA)-induced glucose uptake, purified PKC activity and in vitro 80 kDa protein phosphorylation to elucidate the relationship between insulin-induced PI 3-kinase and PKC activations. Pretreatment with 10(-12)-10(-6) M wortmannin for 60 min resulted in a dose-responsive reduction of 10 nM insulin-stimulated glucose uptake in rat adipocytes. Pretreatment with 10(-6) M wortmannin resulted in 80% and 20% decreases of glucose uptake stimulated by insulin and TPA, respectively. Partially purified rat brain PKC activity and 80 kDa protein in vitro phosphorylation of rat adipocyte cytosol by addition of Ca2+ and phospholipid were dose-dependently decreased by 10(-8)-10(-6) M wortmannin; 20% decrease of PKC activity and 50% decrease of 80 kDa protein phosphorylation by 10(-6) M wortmannin were observed. These results suggest that wortmannin has a potent inhibitory effect on PI 3-kinase and a weak inhibitory effect on PKC activity, and both effects cause a significant inhibition of insulin-stimulated glucose uptake in rat adipocytes.
Ishizuka
T
T
Third Department of Internal Medicine, Gifu University School of Medicine, Japan.
Nagashima
T
T
Yamamoto
M
M
Kajita
K
K
Yamada
K
K
Wada
H
H
Itaya
S
S
Yasuda
K
K
Nozawa
Y
Y
D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; D017667:Adipocytes / Q000187:drug effects* / Q000378:metabolism; D000730:Androstadienes / Q000494:pharmacology*; D000818:Animals; D003847:Deoxyglucose / Q000378:metabolism; D004789:Enzyme Activation; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D005947:Glucose / Q000378:metabolism*; D007328:Insulin / Q000494:pharmacology; D007527:Isoenzymes / Q000187:drug effects / Q000378:metabolism; D007546:Isoquinolines / Q000494:pharmacology; D008297:Male; D019869:Phosphatidylinositol 3-Kinases; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000037:antagonists & inhibitors*; D010879:Piperazines / Q000494:pharmacology; D011493:Protein Kinase C / Q000187:drug effects* / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D013268:Stimulation, Chemical; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D000077191:Wortmannin
D000730:Androstadienes; D004791:Enzyme Inhibitors; D007328:Insulin; D007527:Isoenzymes; D007546:Isoquinolines; D010879:Piperazines; D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; D003847:Deoxyglucose; D019869:Phosphatidylinositol 3-Kinases; D017853:Phosphotransferases (Alcohol Group Acceptor); D011493:Protein Kinase C; D005947:Glucose; D013755:Tetradecanoylphorbol Acetate; D000077191:Wortmannin