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Modification of glutathione S-transferase by selected glycating agents
Průchová, Zuzana ; Dršata, Jaroslav (advisor) ; Trnková, Lucie (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Zuzana Průchová Supervisor: Prof. MUDr. Jaroslav Dršata, CSc. Title of diploma thesis: Modification of Glutathione S-transferase by selected glycating agents Glycation is a non-enzymatic reaction, which leads to the binding of carbonyl compounds to free amino groups of amino acids, peptides, proteins or other biomolecules. This process is closely related to impaired protein function in diabetes mellitus, during aging, and in neurodegenerative diseases. The main purpose of this thesis was to monitor the impact of several glycating agents on the properties of the enzyme glutathione S-transeferase (GST, EC 2.5.1.18) which plays a key role in the detoxification of many exogenous as well as endogenous compounds. Methylglyoxal (0.5-2 mM), glyceraldehyde (0.5-10 mM), glucose (50 mM), and fructose (50 mM) were selected as glycating agents. The course of protein glycation was evaluated by following methods: GST catalytic activity assessment, determination of primary amino groups, assessment of AGEs formation using fluorescence and Western blotting, generation of high molecular cross-links and aggregates by denaturationg polyacrylamide gel electrophoresis (SDS-PAGE), changes in GST molecular...
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