Incorporation and activation of a membrane-bound enzyme in bilayers of liposomes.
The effects of lipid composition and fluidity of lipid bilayers on incorporation and activation of membrane-bound D-fructose dehydrogenase are described in this study. The incorporation of the enzyme into bilayers of small unilamellar vesicles (SUV) made of several phospholipids resulted in enzyme activation with magnitudes higher than that observed in the presence of Triton X-100, indicating that this higher activation is due to lipid-protein interaction. The activity was highest in the presence of SUV formed by the addition of 10% DL-alpha-dipalmitoylphosphatidylethanolamine to L-alpha-dimyristoylphosphatidylcholine, which resulted in eightfold higher activation compared with that of the enzyme in its free state. This activation did not appear to be due to the degree of incorporation of the enzyme, indicating that incorporation is distinct from the activation event. Thus, it is probably the lipid environment that leads to higher activation of the enzyme. A break in the Arrhenius plot of the activity of the membrane-bound enzyme at temperatures close to the phase transition of the phospholipid implies that changes in the physical state of the lipid bilayer influence the enzyme activity. Furthermore, immobilization of D-fructose dehydrogenase, previously adsorbed to SUV, on urethane prepolymer also resulted in about eightfold higher activation than that of the free enzyme.
Kheirolomoom
A
A
Chemical Engineering Department, Kyoto University, Japan.
Miyazato
K
K
Katoh
S
S
Sada
E
E
D002237:Carbohydrate Dehydrogenases / Q000737:chemistry*; D004789:Enzyme Activation; D004795:Enzyme Stability; D004800:Enzymes, Immobilized / Q000737:chemistry*; D007700:Kinetics; D008051:Lipid Bilayers / Q000737:chemistry*; D008081:Liposomes / Q000737:chemistry*; D008560:Membrane Fluidity; D008565:Membrane Proteins / Q000737:chemistry; D010743:Phospholipids / Q000032:analysis
D004800:Enzymes, Immobilized; D008051:Lipid Bilayers; D008081:Liposomes; D008565:Membrane Proteins; D010743:Phospholipids; D002237:Carbohydrate Dehydrogenases