Mechanisms of short-term (second range) regulation of the activities of enzymes of lipid and phospholipid metabolism in secretory cells

• Published in: Advances in enzyme regulation Vol. 28; p. 35
• Main Authors: Söling, H D, Fest, W, Machoczek, K, Schmidt, T, Esselmann, H, Fischer, M
• Format: Journal Article
• Language: English
• Published: England 1989
• Subjects: Acyltransferases - metabolism; Animals; Annexins; Calcium-Binding Proteins - pharmacology; Calcium-Binding Proteins - physiology; Carbachol - pharmacology; Diacylglycerol Kinase; Diacylglycerol O-Acyltransferase; Guinea Pigs; Homeostasis; In Vitro Techniques; Isoproterenol - pharmacology; Kinetics; Lipid Metabolism; Microsomes - enzymology; Parotid Gland - drug effects; Parotid Gland - enzymology; Phospholipases A - antagonists & inhibitors; Phospholipids - metabolism; Phosphotransferases - metabolism; Signal Transduction; Type C Phospholipases - metabolism
• ISSN: 0065-2571
• DOI: 10.1016/0065-2571(89)90062-9

In isolated guinea pig parotid gland lobules the activities of the following enzymes were measured 30 sec after stimulation with either 2 X 10(-5) M isoproterenol or 10(-5) M carbachol: glycerol kinase (EC 2.7.1.30), glycerolphosphate acyltransferase (EC 2.3.1.15), lysophosphatidate acyltransferase (EC 2.3.1.51), phosphatidate phosphohydrolase (EC 3.1.3.4), diacylglycerol acyltransferase (EC 2.3.1.20), diacylglycerol kinase (EC 2.7.1.107), and CDP-diacylglycerol synthetase (EC 2.7.7.41). Lyso-phosphatidate acyltransferase, diacylglycerol kinase, and diacylglycerol acyltransferase exhibited significant increases following stimulation by both types of agonists. Stimulation of the activities of these three enzymes occurred also following in vitro incubation with the catalytic subunit of cAMP-dependent protein kinase or a Ca2+/calmodulin-dependent protein kinase II. These effects could be reversed by incubation with various protein phosphatases. When cells were first stimulated with either type of agonist, subsequent incubation with protein kinases was almost ineffective. Activation by the two types of protein kinases was not additive, indicating that they activate by phosphorylating identical sites on the enzyme proteins. The other enzymes examined showed no or only minor changes and their activities could not be affected by in vitro incubation with the two types of protein kinases. The results explain the rapid changes in acyl-group transfer from acyl-CoA to neutral lipids observed previously during the first seconds after stimulation of guinea pig parotid gland lobules with isoproterenol or carbachol (1). An analysis of a potential role of lipocortins for the regulation of phosphoinositide-specific phospholipases C reveals that these proteins do indeed inhibit these enzymes, but that this inhibition results from a calcium-dependent interaction of the lipocortins with the phospholipid substrate. A physiological role of lipocortins for the regulation of phospholipases is doubtful.