Diacylglycerol lipase and kinase activities in rat brain microvessels.
Diacylglycerols can accumulate transiently in intact cells as a consequence of the degradation of phosphatidylinositol by phospholipase C, but little information is available concerning their metabolic fate in the vascular endothelium. Diacylglycerol lipase and kinase activities were measured in rat brain microvessel preparations. Lipase activity, measured by the release of free fatty acids, was much greater at pH 4.5 than at pH 7. The acid lipase was predominantly particulate and likely originated in lysosomes, whereas the neutral lipase was mainly soluble. The fatty acid at the sn-1 position of the diacylglycerol substrate was hydrolyzed faster than that at the sn-2 position at both pH 4.5 and 7. The 2-monoacylglycerol accumulated at pH 4.5 but not at 7 due to the presence of a monoacylglycerol lipase activity with a neutral pH optimum. The formation of phosphatidic acid (kinase activity) was also measured in microvessels. When lipase and kinase activities were measured simultaneously, the formation of phosphatidic acid from a 1-palmitoyl-2-[1-14C]oleoyl-sn-glycerol substrate was 4-fold greater than the release of fatty acid (oleate) from the sn-2 position. Introduction of arachidonic acid to the sn-2 position of the diacylglycerol substrate increased kinase activity but reduced lipase activity. The release of fatty acids from the sn-2 position of phosphatidic acid could not be detected.
Hee-Cheong
M
M
Fletcher
T
T
Kryski
S K
SK
Severson
D L
DL
D000818:Animals; D001921:Brain / Q000098:blood supply / Q000201:enzymology*; D055598:Chemical Phenomena; D002621:Chemistry; D019852:Diacylglycerol Kinase; D006863:Hydrogen-Ion Concentration; D008071:Lipoprotein Lipase / Q000378:metabolism*; D008833:Microcirculation / Q000201:enzymology; D009131:Muscle, Smooth, Vascular / Q000201:enzymology; D010770:Phosphotransferases / Q000378:metabolism*; D051381:Rats; D013329:Structure-Activity Relationship; D013347:Subcellular Fractions / Q000032:analysis; D013379:Substrate Specificity
D010770:Phosphotransferases; D019852:Diacylglycerol Kinase; D008071:Lipoprotein Lipase