METHOD FOR TRAPPING INTERMEDIATES FORMED BY THE REACTION BETWEEN AN OXIDOREDUCTASE AND ITS SUBSTRATES

摘要

The present invention is the method to trap reliably the reaction intermediates of an oxidoreductase, such as cytochrome-c oxidase. <??>The present invention is a method for trapping reaction intermediates of an oxidoreductase comprising the steps of: (the first step) dissolving an oxidoreductase, a photoinduced reducing agent that releases electrons by light-irradiation, amine-type electron donor and a substrate for said oxidoreductase in water and mixing these; (the second step) cooling the mixture prepared in the first step to 70 SIMILAR 270K to be frozen; (the third step) irradiating the frozen mixture prepared in the second step at 70 SIMILAR 270K with a light in a wavelength region including the absorbing wavelength of said photoinduced reducing agent; and (the fourth step) raising the temperature of the frozen mixture prepared in the third step to the temperature of 80 SIMILAR 270K that is higher than the temperature of the third step.