title: Lipoprotein lipase in cultured heart cells: characteristics and cellular location.
abstract: Lipase activity extracted from cultured neonatal rat heart cells was characterized and identified as lipoprotein lipase. Enzyme activity was stimulated by human apoC-II and rat serum; serum stimulation was prevented by human apoC-I and by apoC-II. Lipolysis was maximal at pH 8.0 and was inhibited by protamine sulfate, NaCl, and high concentrations of heparin. About 50% of heart cell lipase activity applied to heparin-Sepharose bound to the gel and was eluted with a NaCl gradient. A peak of lipase activity was observed at 0.84 M NaCl. Neonatal rat heart cells in culture are a mixture of muscle and non-muscle cells. To determine the cellular location of the lipoprotein lipase, enzyme activity and muscle cell content of the cultures were determined. Myosin ATPase was used as an index of muscle cell content since ATPase specific activity correlated (r = +0.97) with muscle cell content determined immunofluorescently. When muscle cell content of cultures was decreased or increased by differential plating, lipase specific activity was constant. Moreover, lipase specific activity was constant during culture growth despite a decrease in muscle cell content. It was concluded that lipoprotein lipase activity of cultured heart cells is not associated solely with either muscle or non-muslce cells.
mesh_terms: D000251:Adenosine Triphosphatases / Q000378:metabolism; D000818:Animals; D000831:Animals, Newborn; D001053:Apolipoproteins / Q000097:blood / Q000494:pharmacology; D002478:Cells, Cultured; D002846:Chromatography, Affinity; D003470:Culture Media; D005260:Female; D006493:Heparin / Q000494:pharmacology; D008071:Lipoprotein Lipase / Q000302:isolation & purification / Q000378:metabolism*; D008297:Male; D009206:Myocardium / Q000201:enzymology*; D011479:Protamines / Q000494:pharmacology; D051381:Rats
chemical_list: D001053:Apolipoproteins; D003470:Culture Media; D011479:Protamines; D006493:Heparin; D008071:Lipoprotein Lipase; D000251:Adenosine Triphosphatases