Hepatic monoacylglycerol acyltransferase activity in HA1 and HA7 hepatoma/hepatocyte hybrid cells: regulation by insulin and dexamethasone and by cell density.
Hepatic monoacylglycerol acyltransferase (MGAT) (EC 2.3.1.22) is a developmentally-expressed enzyme that catalyzes the stereospecific synthesis of sn-1,2-diacylglycerol from sn-2-monoacylglycerol and long-chain fatty acyl-CoA. In order to study the regulation of MGAT, we developed a rapid assay that can be performed directly on permeabilized HA rat hepatocyte/hepatoma hybrid cells, a line that expresses levels of hepatic MGAT activity and a lipogenic program characteristic of fetal hepatocytes. In permeabilized HA cells, MGAT activity was proportional to the time of incubation and was highly dependent on added sn-2-monoacylglycerol and palmitoyl-CoA. The apparent Km values were 16.6 and 12.7 microM for palmitoyl-CoA and 2-monooleoylglycerol, respectively. Activity was low with the 1(3)- and sn-2-ether analogs of monooleoylglycerol, supporting the conclusion that the cells express the hepatic isoenzyme of MGAT. MGAT activity increased directly with cell density and was unrelated to the number of days in culture. Long-term incubation (2-4 days) of HA cells with various hormones (including triiodothyronine, human placental lactogen, epidermal growth factor, glucagon and growth hormone) showed that only a combination of dexamethasome and insulin resulted in significantly decreased MGAT activity. None of these hormones affected MGAT activity in short-term (0.5-4 h) incubations. These studies suggest that the developmental decline in rat hepatic MGAT activity may be regulated by glucocorticoids and insulin, hormones that increase during and after the second postnatal week.
Coleman
R A
RA
Department of Nutrition, University of North Carolina 27599-7400.
D000217:Acyltransferases / Q000378:metabolism*; D000818:Animals; D002452:Cell Count; D002455:Cell Division; D002463:Cell Membrane Permeability / Q000187:drug effects; D003907:Dexamethasone / Q000494:pharmacology*; D004075:Diglycerides / Q000032:analysis; D004789:Enzyme Activation / Q000187:drug effects; D015870:Gene Expression; D005989:Glycerides / Q000032:analysis; D006822:Hybrid Cells / Q000187:drug effects* / Q000201:enzymology; D007328:Insulin / Q000494:pharmacology*; D008099:Liver / Q000201:enzymology; D010171:Palmitoyl Coenzyme A / Q000032:analysis; D051381:Rats
D004075:Diglycerides; D005989:Glycerides; D007328:Insulin; D010171:Palmitoyl Coenzyme A; D003907:Dexamethasone; C005953:monoolein; D000217:Acyltransferases; C021802:2-acylglycerol O-acyltransferase