Isolation of succinate dehydrogenase isoform from green leaves of maize by ion exchange chromatography

Abstract

Succinate dehydrogenase is an enzyme complex that participates in the course of the catabolic anabolic metabolism and in particular in the tricarboxylic acid cycle, electron transport chain, gluconeogenesis. It is known that plant organisms encountered several forms of the enzyme, particularly in maize and Arabidopsis seeds 4 forms SDH detected. Succinate dehydrogenase structure study is an important task, since the change in the spatial organization of the configuration of the protein molecule ultimately determines its functional properties required of the metabolic fluxes. Poorly understood is the question of the mechanisms of regulation at the level of SDH cell metabolites.
Succinate dehydrogenase isoenzyme spectrum research in green leaves of maize by polyacrylamide gel electrophoresis followed by specific staining for SDH activity showed the presence of two forms with different electrophoretic mobility, probably with different functions in the cell. 5-step purification was carried out to obtain a highly purified preparation of SDH from green leaves of corn. As defining purification steps were carried out ion exchange chromatography, will highlight some of the test forms of the enzyme in a homogeneous state.
Homogeneous preparations of succinate dehydrogenase was used to study their regulatory characteristics. Investigation of the effect of ATP concentration on the activity SDH1 and SDH2 showed difference in their functioning. Both isoforms are inhibited by high ATP concentrations, however, to form SDH1 significant decrease in activity observed at concentrations higher than 50 microM and for form SDH2 - at more than 20 mM. Consequently, we can assume that it is capable of being ATP regulatory component of cells, controlling the rate of oxidative metabolism in the succinate dehydrogenase level in terms of an active photosynthesis.