Isolation of isoenzymes of succinate dehydrogenase from corn seed by ion exchange chromatography

Abstract

Succinate dehydrogenase is a complex enzyme complex that ensures the flow of energy and constructive metabolism. In a number of plant organisms, such as arabidopsis and corn, it is found that there are 4 forms of this enzyme that participate in the cycle of tricarboxylic acids, electron transport chain, gluconeogenesis and amino acids metabolisms. Investigation of the mechanisms of regulation of isoenzymes succinate dehydrogenase, including at the level of metabolites of the cell, is an important task, as it provides a delicate control over the course and redistribution of metabolic cell streams, depending on its needs. Since the change in the spatial organization of the configuration of the protein molecule ultimately determines its functional properties, the necessary implementation of metabolic fluxes.

The study of the isoenzyme spectrum of succinate dehydrogenase in corn seed shields by the method of polyacrylamide gel electrophoresis followed by specific staining for SDH activity showed the presence of four forms with different electrophoretic mobility, probably performing different functions in the cell. To obtain a highly purified SDH preparation from the corn seed shields, a 4-step purification was carried out. As a determining purification stage, ion-exchange chromatography was carried out, which made it possible to separate the individual forms of the enzyme under study in an electrophoretically homogeneous state. The preparations of isoforms of succinate dehydrogenase obtained in the homogeneous state allow further study of their regulatory and kinetic properties, with the aim of establishing mechanisms of regulation of the intensity of oxidative and constructive metabolism at the level of succinate dehydrogenase