Glycolipids isolated from cultured rat hepatocytes: analysis of their role in insulin signal transduction.
The precise mechanism by which insulin elicits its effects remains to be fully determined. A glycophospholipid, isolated from H35 cells, has been proposed as a possible precursor for an insulin-generated second messenger that mediates the intracellular effects of insulin. This glycolipid contains a hexosamine moiety, inositol, galactose and palmitate. We have isolated a glycolipid from cultured rat hepatocytes that exhibits chromatographic and radiolabelling characteristics similar to this proposed precursor. The glycolipid can be radiolabelled with glucosamine, galactosamine, galactose and palmitate, but not myristate or myoinositol. Incorporation of radiolabel into this glycolipid was insensitive to the presence of either insulin (10(-7) M) or phosphatidylinositol-specific phospholipase C (PI-PLC) in the culture medium. The cultured hepatocytes used exhibited normal insulin responses with respect to glycogen turnover and gene expression. Treatment of partially purified glycolipid with either PI-PLC or nitrous acid did not result in the generation of an aqueous soluble phosphooligosaccharide indicating that the glycolipid was not cleaved by either agent. This is in contrast to the reported cleavage of the glycolipids found in H35 hepatoma and lymphocytes. These results question the role of the putative phosphooligosaccharide mediator in the intracellular transduction system activated by insulin.
Department of Biochemistry and Molecular Biology, University of Manchester, U.K.
A J
AJ
Whatmore
A J
AJ
Bate
A J
AJ
Dickson
D000418:Albumins / Q000096:biosynthesis; D000818:Animals; D002478:Cells, Cultured; D000242:Cyclic AMP / Q000031:analogs & derivatives / Q000494:pharmacology; D003907:Dexamethasone / Q000494:pharmacology; D005786:Gene Expression Regulation / Q000187:drug effects; D006017:Glycolipids / Q000302:isolation & purification / Q000494:pharmacology / Q000502:physiology*; D007328:Insulin / Q000494:pharmacology / Q000502:physiology*; D008099:Liver / Q000737:chemistry* / Q000166:cytology; D008112:Liver Glycogen / Q000378:metabolism; D043265:Phosphatidylinositol Diacylglycerol-Lyase; D010729:Phosphoenolpyruvate Carboxykinase (GTP) / Q000096:biosynthesis; D054801:Phosphoinositide Phospholipase C; D010727:Phosphoric Diester Hydrolases / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D015398:Signal Transduction / Q000502:physiology*; D013873:Thionucleotides / Q000494:pharmacology
D000418:Albumins; D006017:Glycolipids; D007328:Insulin; D008112:Liver Glycogen; D013873:Thionucleotides; C015929:8-((4-chlorophenyl)thio)cyclic-3',5'-AMP; D003907:Dexamethasone; D000242:Cyclic AMP; D010727:Phosphoric Diester Hydrolases; D054801:Phosphoinositide Phospholipase C; D010729:Phosphoenolpyruvate Carboxykinase (GTP); D043265:Phosphatidylinositol Diacylglycerol-Lyase