Characterization of the interaction of phospholipase A(2) with phosphatidylcholine-phosphatidylglycerol mixed lipids.
The first requirement in the hydrolysis of phospholipid bilayers by phospholipase A(2) is the interaction of the enzyme with the bilayer surface. The catalytic ability of phospholipase A(2) has been shown to be extremely sensitive to the topology of the bilayer to which it binds and hydrolyzes. Phospholipid bilayer properties and composition such as unsaturation, charge, and the presence of reaction products are known regulators of the catalytic activity of phospholipase A(2) toward the phospholipids and influences the binding of enzyme to the membrane. We show in this paper that the effect of increased anionic lipid results in enhanced binding that can be described quantitatively in terms of a simple phenomenological model. However, the interaction with anionic lipid does not singularly dominate the thermodynamics of binding, nor can the lag phase observed in the time course of hydrolysis of large unilamellar vesicles simply be the result of limited interaction between the enzyme and the bilayer. Furthermore, we show that phospholipase A(2) from Akgistrodon piscivorus piscivorus can exist in at least two bilayer-bound states and that the absence of a fluorescence change upon mixing the enzyme with lipid bilayers does not necessarily indicate the absence of an interaction.
Gadd
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ME
Departments of Biochemistry and Molecular Genetics and Pharmacology, University of Virginia, Charlottesville, Virginia 22908, USA. mgadd@emory.edu
Biltonen
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RL
D003435:Crotalid Venoms / Q000201:enzymology; D008051:Lipid Bilayers / Q000737:chemistry* / Q000378:metabolism*; D008956:Models, Chemical; D008968:Molecular Conformation; D010713:Phosphatidylcholines / Q000378:metabolism*; D010715:Phosphatidylglycerols / Q000378:metabolism*; D010741:Phospholipases A / Q000378:metabolism*; D011485:Protein Binding
D003435:Crotalid Venoms; D008051:Lipid Bilayers; D010713:Phosphatidylcholines; D010715:Phosphatidylglycerols; D010741:Phospholipases A