Isolation of isoenzymes of succinate dehydrogenase from pea leaves by ion exchange chromatography

Abstract

Succinate dehydrogenase (SDH) is a multifunctional enzyme and for this reason has an almost universal distribution among living organisms. This enzyme plays a key role in the regulation of aerobic respiration. In addition, SDH is a membrane-bound enzyme, which determines its polyfunctionality - participation in the functioning of the TCA and the operation of the electron transport chain (complex II). Despite the large amount of experimental material on the physicochemical and regulatory characteristics of succinate dehydrogenase from objects of various origins, methods of regulating the activity of this enzyme system in plants under conditions of changing environmental factors remain unknown or poorly studied.

The preparation of highly purified SDH preparations from the leaves was done by a 4-stage purification method. An important stage, which allowed to separate the separate forms of the enzyme, was the stage of ion-exchange chromatography. Desorption with a linear gradient of potassium chloride within the range of 50-125 mM made it possible to separate the individual forms of the enzyme under study in a highly purified state. The obtained preparations of succinate dehydrogenase isoforms will allow further study of their regulatory and kinetic properties in order to establish mechanisms of regulation of the intensity of oxidative and constructive metabolism at the level of succinate dehydrogenase.