Lipolytic stimulation modulates the subcellular distribution of hormone-sensitive lipase in 3T3-L1 cells.
3T3-L1 cells have been a useful model system for studying adipocyte differentiation and metabolism. They acquire a hormone-sensitive lipase during differentiation (Kawamura, M., et al. 1981. Proc. Natl. Acad. Sci. USA. 78: 732-735). In the present study the control of lipolysis in these cells was investigated. Basal glycerol release from cell monolayers was 437 nmol/mg protein per hr, and could be stimulated approximately 6-fold by exposure to 1 microM isoproterenol. Subcellular fractionation of stimulated cells revealed a redistribution of triglyceride lipase activity: loss from the infranatant fraction and increase in the pellet fraction. The redistribution was dosage-dependent and reversible. Treatment of intact cells with 8-bromoadenosine 3':5' cyclic monophosphate elicited similar redistribution of the lipase activity; however, disruption and incubation of untreated cells in the presence of ATP and either cyclic AMP or the catalytic subunit from cAMP-dependent protein kinase did not. The lipase activity in the pellet fraction was increased 3- to 4-fold after maximal lipolytic stimulation of intact cells, whereas phosphorylation of the enzyme in vitro yielded 1.4- to 1.6-fold stimulation in all subcellular fractions from untreated cells. The lipase found in the particulate fraction has the same properties as the previously characterized infranatant enzyme. It is suggested that interaction of the lipase with substrate and associated intracellular membranes may be a novel feature of the regulation of lipolysis.
D015056:1-Methyl-3-isobutylxanthine / Q000494:pharmacology; D015124:8-Bromo Cyclic Adenosine Monophosphate / Q000494:pharmacology; D000273:Adipose Tissue / Q000201:enzymology / Q000648:ultrastructure*; D000324:Adrenocorticotropic Hormone / Q000494:pharmacology; D000818:Animals; D002460:Cell Line; D004305:Dose-Response Relationship, Drug; D007545:Isoproterenol / Q000494:pharmacology; D008066:Lipolysis; D008071:Lipoprotein Lipase / Q000378:metabolism; D051379:Mice; D002787:Sterol Esterase / Q000378:metabolism*; D013347:Subcellular Fractions / Q000201:enzymology*; D014018:Tissue Distribution
D015124:8-Bromo Cyclic Adenosine Monophosphate; D000324:Adrenocorticotropic Hormone; D002787:Sterol Esterase; D008071:Lipoprotein Lipase; D007545:Isoproterenol; D015056:1-Methyl-3-isobutylxanthine