The activity of glutathione peroxidase at liver function disturbance and enzyme isolation by chromatographic methods for the regulatory properties study

Abstract

The aim of the work was to study the activity of glutathione peroxidase (GP, EC 1.11.1.9.) under
liver functions disturbance, as well as the enzyme purification by chromatographic methods for further study
of regulatory properties. It has been found that side by side with objective clinical signs of liver diseases,
marker biochemical and immune parameters changes, there was an increase in the activity of the GP in the
blood serum of patients with non-alcoholic steatohepatitis, developing under type 2 diabetes, and druginduced
hepatitis, emerged on the background of anti- tuberculosis chemotherapy, and also in rat blood serum
and liver at experimental toxic hepatitis (ETG) compared to the control level. The purification of GP
from rat liver in normal conditions and ETG by homogenization of the material, gel-filtration on Sephadex
G-25, ion-exchange chromatography on DEAE-cellulose, concentration of the enzyme fractions by ultrafiltration
on Amicon cell, gel-chromatography on Toyopearl HW-65 has been carried out. As a result, the enzyme
preparations with a specific activity of 1.46 and 3.64 E/ protein mg, yield 7.2 and 8.0% respectively
have been obtained. Further they were used for the study of GP regulatory properties. Based on the received
results we can assume that one of the mechanisms associated with the increase of the enzyme activity under liver functions disturbance may be linked with conformational restructurings of the molecule under conditions
of enhanced free radicals generation.