Engineering of functional supramacromolecular complexes of proteins (enzymes) using reversed micelles as matrix microreactors |
| |
Authors: | Kabanov Alexander V; Klyachko Natalya L; Nametkin Sergei N; Merker Steffen; Zaroza Anna V; Bunik Vita I; Ivanov Mikhail V; Levashov Andrey V |
| |
Affiliation: | Department of Chemical Enzymology, Faculty of Chemistry, M.V. Lomonosov Moscow State University 119899, GSP, USSR
1Department of Biochemistry, Faculty of Biology, M.V. Lomonosov Moscow State University Moscow, 119899, GSP, USSR
2Present address: Research Centre of Biotechnology Alt Stralau 62, Berlin, 1017, Germany
3A.N. Belozersky Laboratory of Molecular Biology and Bioorganic Chemistry, M.V. Lomonosov Moscow State University Moscow, 119899, GSP, USSR |
| |
Abstract: | The size of the inner water cavity of reversed micelles formedin a triple system water-surfactant-organic solventcan be widely varied by changing the degree of surfactant hydration.This gives grounds to use reversed micelles as matrix microreactorsfor the design of supramolecular complexes of proteins. Usingultracentrifugation analysis, it has been demonstrated thatthe oligomeric composition of various enzymes (ketoglutaratedehydrogenase, alkaline phosphatase, lactic dehydrogenase, glyceraldehyde-3-phosphatedehydrogenase) solubilized in reversed micelles of Aerosol OTsodium bis(2-ethylehexyl)sulfosuccinate] in octane changesupon variation of the degree of hydration. An oligomeric complexforms under conditions when the radius of the micelle innercavity is big enough to incorporate this complex as a whole.At lower degrees of hydration the micelles uncouplesuch complexes to their components. The catalytic propertiesof various oligomeric complexes have been studied. Possibilitiesof using reversed micelles for the separation of subunits ofoligomeric enzymes under non-denaturating conditions have beendemonstrated. In particular, the isolated subunits of alkalinephosphatase, lactic dehydrogenase and glyceralde-hyde-3-phosphatedehydrogenase have been found to be active in Aerosol OT reversedmicelles. The dependences of the catalytic activity of oligomericenzymes represent saw-like curves. The maxima of the catalyticactivity observed at these curves relate to the functioningof various oligomeric forms of an enzyme. The radii of the micelleinner cavity under conditions when these maxima are observedcorrelate with the linear dimensions of the enzyme oligomericforms. Correlation of the position of a maximum with the shapeof an oligomeric complex is discussed. |
| |
Keywords: | alkaline phosphatase/ glyceraldehyde-3-phosphate dehydrogenase/ ketoglutarate dehydrogenase/ lactic dehydrogenase/ reversed micelles |
本文献已被 Oxford 等数据库收录! |
|