Regulation of the catalytic activity and oligomeric composition of enzymes in reversed micelles of surfactants in organic solvents.
The phenomenon of regulation of the catalytic activity of enzymes via changing their oligomeric composition in the system of reversed micelles of sodium bis(2-ethylhexyl)sulfosuccinate (AOT) in octane was studied using alpha-chymotrypsin (CT) from bovine brain and alkaline phosphatase (AP) from calf intestinal mucosa. The dependences of the enzyme catalytic activity on the AOT hydration degree (Wo = [H2O]/[AOT]), the parameter determining the radius (rc) of the inner cavity of micelles, usually represent the bell-shaped curves. The maximal catalytic activity is observed at such Wo when rc is equal to the size of the enzyme molecule. The position of this maximum strictly correlates with the enzyme oligomeric composition. Thus, in the case of CT this is observed at Wo = 12 when rc is equal to the radius (rp) of the CT globule. In the case of artificially produced conjugate containing six cross-linked CT molecules, this is observed at Wo = 43 when rc is equal to the radius of the sphere surrounding the absolute octahedron composed of six CT globules. The dependence of the catalytic activity of AP on Wo represents a curve with two maxima that are observed when rc is equal to rp of either AP monomer (Wo = 17) or AP dimer (Wo = 25). Ultracentrifugation experiments revealed that variation of Wo causes a change in the oligomeric composition of AP - its transition from monomeric (Wo less than 20) to dimeric form (Wo greater than 20). Hence, the observed maxima correspond to functioning of different oligomeric forms of AP.
Kabanov
A A
AA
Department of Chemical Enzymology, Moscow State University, USSR.
Nametkin
S N
SN
Klyachko
N L
NL
Levashov
A V
AV
D000469:Alkaline Phosphatase / Q000737:chemistry / Q000378:metabolism*; D000818:Animals; D002384:Catalysis; D002417:Cattle; D002918:Chymotrypsin / Q000737:chemistry / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D007413:Intestinal Mucosa / Q000201:enzymology; D046911:Macromolecular Substances; D008823:Micelles; D015394:Molecular Structure; D009795:Octanes; D010179:Pancreas / Q000201:enzymology; D014867:Water
D046911:Macromolecular Substances; D008823:Micelles; D009795:Octanes; D014867:Water; D000469:Alkaline Phosphatase; D002918:Chymotrypsin; C026728:octane