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Title

Μελέτη του ΑΤΡ ως υπόστρωμα της 15-Λιποξυγενάσης-1

Alternative Title

Study of ATP as a substrate of 15-Lipoxygenase-1

Author

Βραχνά, Μαρία

Thesis advisor

Ελευθεριάδης, Νικόλαος

Abstract

15-Lipoxygenase-1 (15-LOX-1) is a key enzyme in mammalian cells, mainly involved in lipid metabolism. It is located in the cytoplasm of immune and epithelial cells and targets polyunsaturated fatty acids such as arachidonic and linoleic acids. By catalyzing the oxygenation of fatty acids at the 15th carbon position, it produces hydroperoxyacetate tetraenoic acid derivatives (HPETE), which serve as precursors such as lipid bioactive mediators. The importance of 15- LOX1 extends to various physiological and pathological processes, including inflammation, atherosclerosis, cancer and neurodegenerative diseases. Overall, 15-lipoxygenase-1 is characterized as a multifaceted, important enzyme for lipid metabolism and cellular physiology, thus making it a prime target for therapeutic interventions in different diseases. In this thesis, the heterologous expression and purification process of the enzyme is first presented and then the ability to hydrolyze ATP by ADT and inorganic phosphorus is examined. After the hydrolysis of ATP by the enzyme was established and quantified, factors likely to affect the rate of hydrolysis were examined, such as different concentrations of ATP and enzyme, reaction incubation time and thermal incubation of the enzyme at different temperatures. Then, the inhibition of catalytic ATP hydrolysis using two inhibitors and the dependence of ATP hydrolysis under the presence of substrate were investigated. Finally, the ATP hydrolysis reaction of the enzyme under the coexistence of both the enzyme substrate and a synergistic inhibitor was studied.

Language

Greek

Subject

ATP hydrolysis

ATP quantification

Factors affecting ATP hydrolysis

Heterologous expression

Protein purification

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