Metabolism of dioctanoylglycerol by isolated cardiac myocytes.
Diacylglycerol (DG) metabolism by intact cardiac myocytes isolated from adult rat hearts and by broken cell preparations from myocytes was investigated in experiments with [3H]dioctanoylglycerol (diC8), a cell-permeable diacylglycerol analog. In a low-speed supernatant fraction, the Km and Vmax for DG kinase activity (formation of phosphatidic acid) was 22 microM and 110 nmol/h/mg, respectively, whereas the Km and Vmax for DG lipase activity (formation of monoacylglycerol) was 80 microM and 1000 nmol/h/mg. At a substrate concentration of 80 microM diC8, lipase activity was 7-fold greater than kinase activity. The majority of DG kinase activity was recovered in the soluble subcellular fraction; DG lipase activity was localized in a microsomal fraction. When [3H]diC8 was incubated with intact cardiac myocytes, 10-fold more radioactivity was incorporated into the products of the lipase pathway (monoacylglycerol and free glycerol) as compared to incorporation into the total phospholipid fraction which contained phosphatidic acid. This predominance of metabolism by hydrolysis through the lipase pathway was observed consistently when the incubation time, content of cardiac myocytes and concentration of exogenous diC8 was varied. Therefore, results from both in vitro determinations of enzyme activities in broken cell preparations and flux studies with intact cells have indicated that the lipase pathway is the principal enzymatic mechanism for the metabolism of diC8 in cardiac myocytes.
Hee-Cheong
M
M
Department of Pharmacology and Therapeutics, Faculty of Medicine, University of Calgary, Alberta, Canada.
Severson
D L
DL
D000818:Animals; D019852:Diacylglycerol Kinase; D004075:Diglycerides / Q000378:metabolism*; D005989:Glycerides / Q000378:metabolism*; D066298:In Vitro Techniques; D007700:Kinetics; D008071:Lipoprotein Lipase / Q000378:metabolism; D009206:Myocardium / Q000378:metabolism*; D010770:Phosphotransferases / Q000378:metabolism; D011865:Radioisotope Dilution Technique; D051381:Rats; D013347:Subcellular Fractions / Q000378:metabolism; D014316:Tritium
D004075:Diglycerides; D005989:Glycerides; D014316:Tritium; C044295:1,2-dioctanoylglycerol; D010770:Phosphotransferases; D019852:Diacylglycerol Kinase; D008071:Lipoprotein Lipase