Multiple Factors Influence the Binding of a Soluble, Ca2+-Independent, Diacylglycerol Kinase to Unilamellar Phosphoglyceride Vesicles
- 20 February 1999
- journal article
- research article
- Published by American Chemical Society (ACS) in Biochemistry
- Vol. 38 (11), 3310-3319
- https://doi.org/10.1021/bi982566u
Abstract
We studied the influence of membrane lipids, MgCl2, and ATP on the ability of a soluble diacylglycerol kinase to bind to 100-nm lipid vesicles. The enzyme did not bind detectably to vesicles that contained phosphatidylcholine alone or to vesicles that contained 50 mol % phosphatidylcholine + 50 mol % phosphatidylethanolamine. But it did bind to vesicles that contained anionic phosphoglycerides, and maximal binding occurred (in the presence of MgCl2) when the vesicles contained anionic phosphoglycerides alone. When increasing amounts of phosphatidylcholine were included in phosphatidylserine-containing vesicles, enzyme binding to the vesicles decreased by as much as 1000-fold. However, when increasing amounts of phosphatidylethanolamine were included in phosphatidylserine-containing vesicles, little change in binding occurred until the concentration of phosphatidylserine was reduced to below 25 mol %. These results and results obtained with vesicles that contained various mixtures of anionic phosphoglycerides, phosphatidylcholine, phosphatidylethanolamine, and unesterified cholesterol provided evidence that anionic phosphoglycerides were positive effectors of binding, phosphatidylcholine was a negative effector, and phosphatidylethanolamine and unesterified cholesterol were essentially neutral diluents. Other experiments showed that diacylglycerol and some of its structural analogues also were important, positive effectors of enzyme binding and that addition of ATP to the medium increased their effects. The combined results of the study suggest that the enzyme may bind to vesicles via at least two types of binding sites: one type that requires anionic phospholipids and is enhanced by Mg2+ but inhibited by phosphatidylcholine, and one type that requires diacylglycerol and is enhanced by ATP.Keywords
This publication has 17 references indexed in Scilit:
- Diacylglycerol kinaseThe International Journal of Biochemistry & Cell Biology, 1997
- Cloning of a Novel Human Diacylglycerol Kinase (DGKθ) Containing Three Cysteine-rich Domains, a Proline-rich Region, and a Pleckstrin Homology Domain with an Overlapping Ras-associating DomainJournal of Biological Chemistry, 1997
- Cloning and Characterization of a Glucocorticoid-induced Diacylglycerol KinaseJournal of Biological Chemistry, 1996
- The phase behavior of mixed aqueous dispersions of dipalmitoyl derivatives of phosphatidylcholine and diacylglycerolBiophysical Journal, 1994
- Characterization of Two Cytosolic Diacylglycerol Kinase FormsJournal of Neurochemistry, 1993
- Gene cloning, sequence, expression and in situ localization of 80 kDa diacylglycerol kinase specific to oligodendrocyte of rat brainMolecular Brain Research, 1992
- Purification, cDNA‐cloning and expression of human diacylglycerol kinaseFEBS Letters, 1990
- Different effects of sphingosine, R59022 and anionic amphiphiles on two diacylglycerol kinase isozymes purified from porcine thymus cytosolFEBS Letters, 1989
- Miscibility, chain packing, and hydration of 1-palmitoyl-2-oleoyl phosphatidylcholine and other lipids in surface phasesBiophysical Journal, 1985
- A Rapid and Simple Method for the Determination of Esterified Fatty Acids and for Total Fatty Acids in BloodJournal of Clinical Pathology, 1953