Coordinate regulation of triacylglycerol synthesis and glucose transport by acylation-stimulating protein.
Acylation-stimulating protein (ASP) is the most potent recognized stimulant of triacylglycerol synthesis in human adipocytes. However, its mechanism(s) of action have not yet been elucidated in detail. The present study examines the effects of ASP on membrane transport of glucose and fatty acids in cultured human skin fibroblasts. The data demonstrate that ASP stimulated carrier-mediated glucose transport in a time- and concentration-dependent manner, an effect that was greater than and independent of that observed with insulin. ASP increased the Vmax for glucose transport with no change in the transport Km, suggesting that ASP might result in increased sugar transporters in the plasma membrane. This finding was supported by quantitative Western blot analyses using a monoclonal antibody (G3) that demonstrated an increase in the plasma membrane content of the glucose transporter (Glut 1) with a concomitant decrease in the glucose transporter content of an intracellular membrane fraction. By contrast, ASP had no effect on specific membrane transport of fatty acids. Maximal effects of ASP on triacylglycerol synthesis were demonstrated at saturating levels of both glucose and oleate. Comparable stimulation by ASP in the absence of glucose (with or without pyruvate) was also demonstrated, although the absolute rates of triacylglycerol synthesis were substantially lower. Finally, it was shown that ASP increased the apparent Vmax for triglyceride synthesis without changing its Km. Since ASP will act in the absence of extracellular glucose, ASP must have additional actions, independent of its effect on specific membrane transport of glucose by which it accelerates intracellular triglyceride synthesis.(ABSTRACT TRUNCATED AT 250 WORDS)
Lady Davis Institute, Sir Mortimer B. Davis, Jewish General Hospital, Department of Medicine, McGill University, Montreal, Quebec, Canada.
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Germinario
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Sniderman
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Manuel
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Lefebvre
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Baldo
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Cianflone
D001692:Biological Transport / Q000187:drug effects; D001798:Blood Proteins / Q000494:pharmacology*; D002478:Cells, Cultured; D015926:Complement C3a / Q000031:analogs & derivatives*; D003847:Deoxyglucose / Q000378:metabolism; D005227:Fatty Acids / Q000378:metabolism; D005347:Fibroblasts / Q000378:metabolism; D005947:Glucose / Q000378:metabolism* / Q000494:pharmacology; D006801:Humans; D007328:Insulin / Q000494:pharmacology; D019301:Oleic Acid; D009829:Oleic Acids / Q000494:pharmacology; D014280:Triglycerides / Q000096:biosynthesis*
D001798:Blood Proteins; D005227:Fatty Acids; D007328:Insulin; D009829:Oleic Acids; D014280:Triglycerides; C044541:complement C3a, des-Arg-(77)-; D019301:Oleic Acid; D015926:Complement C3a; D003847:Deoxyglucose; D005947:Glucose