A new strategy for the study of oligomeric enzymes: gamma-glutamyltransferase in reversed micelles of surfactants in organic solvents.
A heterodimeric enzyme (gamma-glutamyltransferase) was studied in the reversed micellar medium of Aerosol OT (AOT) in octane. As was shown earlier, the size (radius) of inner cavity of the AOT-reversed micelles is determined by their hydration degree, i.e., [H2O]/[AOT] molar ratio, in the system. Owing to this, the dependence of hydrolytic, transpeptidation and autotranspeptidation activities of the enzyme on the hydration degree was investigated using L- and D-isomers of gamma-glutamyl(3-carboxy-4-nitro)anilide and glycylglycine as substrates. For all of the reaction types, the observed dependences are curves with three optima. The optima are found at the hydration degrees, [H2O]/[AOT] = 11, 17 and 26 when the inner cavity radii of reversed micelles are equal to the size of light (Mr 21,000) and heavy (Mr 54,000) subunits of gamma-glutamyltransferase, and to their dimer (Mr 75,000), respectively. Ultracentrifugation experiments showed that a change of the hydration degree resulted in a reversible dissociation of the enzyme to light and heavy subunits. The separation of light and heavy subunits of gamma-glutamyltransferase formed in reversed micelles was carried out and their catalytic properties were studied. The two subunits catalyze hydrolysis and transpeptidation reactions; autotranspeptidation reaction is detected only in the case of the heavy subunit. These findings imply that the reversed micelles of surfactants in organic solvents function as the matrices with adjustable size permitting to regulate the supramolecular structure and the catalytic activity of oligomeric enzymes.
Kabanov
A V
AV
Department of Chemistry, Moscow State University, U.S.S.R.
Nametkin
S N
SN
Evtushenko
G N
GN
Chernov
N N
NN
Klyachko
N L
NL
Levashov
A V
AV
Martinek
K
K
D002384:Catalysis; D002498:Centrifugation; D003102:Colloids; D004143:Dioctyl Sulfosuccinic Acid; D008823:Micelles; D008956:Models, Chemical; D009097:Multienzyme Complexes; D009795:Octanes; D012997:Solvents; D013501:Surface-Active Agents; D005723:gamma-Glutamyltransferase / Q000032:analysis / Q000378:metabolism*
D003102:Colloids; D008823:Micelles; D009097:Multienzyme Complexes; D009795:Octanes; D012997:Solvents; D013501:Surface-Active Agents; D004143:Dioctyl Sulfosuccinic Acid; D005723:gamma-Glutamyltransferase; C026728:octane