Microsomal diacylglycerol acyltransferase in rat parotid and submandibular glands: Acylation of 1,2-Dioleoyl-sn-glycerol dispersed with phospholipids

In order to investigate microsomal diacylglycerol acyltransferase activity, ethanol or several detergents have been used as a dispersing agent for water-insoluble substrates. However, ethanol acyltransferase interferes with the activity of this enzyme, and detergents inhibit it. We examined the properties of microsomal diacylglycerol acyltransferase in rat salivary glands without detergents or organic solvents. 1,2-Dioleoyl-sn-glycerol (1,2-diolein) was dispersed by sonication. The activity was measured as the formation rate of [¹⁴C]triglyceride using [1-¹⁴C]palmitoyl-CoA as an acyl-donor. The reaction was dependent on the microsomal protein and 1,2-diolein at least up to 145 μg/ml and 3.6 mM, respectively. The specific activities were 3.91 ± 0.57 and 3.80 ± 0.77 nmol/min per mg protein (SEM, n = 4) in the parotid and submandibular glands, respectively. They were 12- to 20-fold higher than the activities in liver, brain and spleen, and two orders of magnitude higher than that assayed with microsomal endogenous diacylglycerol. Adding tissue phospholipids to 1,2-diolein suspension reduced the concentration of 1,2-diolein required for the maximal velocity. A similar, but reduced, effect was induced by egg yolk phosphatidylcholine in plate of the tissue phospholipids. The level of activity was recovered by adding another phospholipid class to the phosphatidylcholine. The results suggested that the physical condition of the substrate diacylglycerol affects diacylglycerol acyltransferase activity in rat salivary gland microsomes.