A novel approach to study of action of water-insoluble inhibitors of enzymic reactions.
The effect of water-insoluble compounds on enzyme catalytic properties was studied using a colloidal solution of water in organic solvent as reaction medium. In this microheterogeneous medium enzyme is entrapped into hydrated reversed micelles of a surfactant, the dimensions of the internal hole of the micelles being dependent on the ratio of water to surfactant. At sufficiently low values of this ratio the molecule of entrapped enzyme has limited mobility in the micelle. Because of this the interaction of the enzyme with water-insoluble compound which is added in assay solution and intercalated in the surface layer of the micelle may be manifested. The suggested method was used to study the inhibitory action of dihydroriboflavin esters on D-amino acid oxidase from pig kidney and soybean lipoxygenase. The reaction medium was hydrated reversed micelles of Aerosol OT in octane. The method of sedimentation in an analytical ultracentrifuge has shown the dihydroriboflavin esters to be completely included into reversed micelles.
B I
BI
Kurganov
L G
LG
Tsetlin
E A
EA
Malakhova
N A
NA
Chebotareva
V Z
VZ
Lankin
G D
GD
Glebova
V M
VM
Berezovsky
A V
AV
Levashov
K
K
Martinek
D000818:Animals; D003605:D-Amino-Acid Oxidase / Q000037:antagonists & inhibitors; D004143:Dioctyl Sulfosuccinic Acid; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D007668:Kidney / Q000201:enzymology; D016859:Lipoxygenase Inhibitors; D008433:Mathematics; D008823:Micelles; D012256:Riboflavin / Q000494:pharmacology; D012995:Solubility; D013025:Glycine max / Q000201:enzymology; D013552:Swine; D014461:Ultracentrifugation; D014867:Water
D004791:Enzyme Inhibitors; D016859:Lipoxygenase Inhibitors; D008823:Micelles; D014867:Water; D004143:Dioctyl Sulfosuccinic Acid; D003605:D-Amino-Acid Oxidase; D012256:Riboflavin