Molecular details of the activation of soluble phospholipase A2 on lipid bilayers. Comparison of computer simulations with experimental results.
The initial rate of hydrolysis of large unilamellar vesicles of dipalmitoylphosphatidylcholine by phospholipase A2 from the venom of Agkistrodon piscivorus piscivorus is small and elevates gradually until it suddenly increases by a factor of 10 to 1000 depending on the experimental conditions. This abrupt onset of high enzyme activity appears to be correlated to a specific mole fraction of reaction product at which point a cooperative compositional phase transition in the bilayer occurs. Five models that describe the activation process in terms of its being coupled to the putative product-induced lipid transition are presented. These models include one in which the lipid structure enhances the affinity of enzyme binding to the bilayer surface, two in which the equilibrium position between an active and an inactive form of the enzyme-substrate complex is altered, and two in which the rate of a quasi-irreversible spontaneous activation process is increased. Whether the active form of the enzyme is a monomer or dimer is also considered in the last two pairs of models. Computer simulations of time courses for the different models show how a set of four experimental observables distinguishes qualitatively among them. Comparison of the experimental behavior with the computer-simulated behavior of the observables for each model indicates that activation of phospholipase A2 on the lipid surface involves formation of an enzyme dimer which spontaneously converts to an active form. The active enzyme persists in the active state as it exchanges between vesicles. This model of activation is similar to that proposed previously for activation of porcine pancreatic phospholipase A2.
Bell
J D
JD
Department of Zoology, Brigham Young University, Provo, Utah 84602.
Biltonen
R L
RL
D003198:Computer Simulation; D004789:Enzyme Activation; D006868:Hydrolysis; D007700:Kinetics; D008051:Lipid Bilayers; D008244:Lysophosphatidylcholines / Q000494:pharmacology; D008962:Models, Theoretical; D010741:Phospholipases A / Q000378:metabolism*; D054467:Phospholipases A2; D013050:Spectrometry, Fluorescence
D008051:Lipid Bilayers; D008244:Lysophosphatidylcholines; D010741:Phospholipases A; D054467:Phospholipases A2