Impact of Effectors on the Catalytic Activity of Galactonolactone Oxidase from Trypanosoma cruzi

Chudin, A.A.; Kudryashova, E.V.

Авторы: Chudin A.A., Kudryashova E.V.
Журнал: Russian Journal of Bioorganic Chemistry
Том: 50
Номер: 4
Год издания: 2024
Издательство: Maik Nauka/Interperiodica Publishing
Местоположение издательства: Russian Federation
Первая страница: 1294
Последняя страница: 1305
DOI: 10.1134/S1068162024040071
Аннотация: Abstract—Objective: Galactonolactone oxidase from Trypanosoma cruzi is a membrane enzyme catalyzingthe final step of the biosynthesis of vitamin C, an antioxidant essential for T. cruzi causing Chagas disease. So, galactonolactone oxidase can be considered as a promising drug target. This work devoted to investigation of impact of the effectors, 1,4-benzoquinone, coenzymes Q, and their structural analogs, on the activity of galactonolactone oxidase from Trypanosoma cruzi and of the model homologous enzyme L-galactono-1,4-lactone dehydrogenase from Arabidopsis thaliana. Methods: Recombinant expression of galactonolactone oxidase and L-galactono-1,4-lactone dehydrogenase was carried out in Escherichia coli and refolding of galactonolactone oxidase was performed using reverse micelles system. Spectrophotometric determination of the activity of both enzymes was carried out in a surfactant reverse micelles system using a combination of one of the potentialeffectors and 2,6-dichlorophenolindophenol, a dye, which becomes colorless upon interaction with the product of reaction. Results and Discussion: Using two forms of L-galactono-1,4-lactone dehydrogenase, wild-type (dehydrogenase) and mutant (exhibiting oxidase activity), the role of 1,4-benzoquinone and its analogs as electron acceptors of galactono-1,4-lactone dehydrogenase and galactonolactone oxidase was revealed. It was established that compounds containing methoxy groups (coenzyme Q0, 2,6-dimethoxy-1,4-benzoquinone) are more effective electron acceptors for galactonolactone oxidase compared to those lacking OCH3 groups (2,5-dihydroxy-1,4-benzoquinone). Conclusions: Basing on the use of 2,6-dimethoxy-1,4-benzoquinone as an electron acceptor, an approach was proposed to the spectrophotometric measurement of the galactonolactone oxidase activity by the changes in the absorption of the electron acceptor in the absence of additional components (a dye that becomes colorless when interacting with the reaction product, ascorbate). The results obtained enable more targeted search for inhibitors of galactonolactone oxidase, which may be regarded as a basis for the development of selective drugsagainst Chagas disease caused by T. cruzi.
Добавил в систему: Чудин Андрей Алексеевич

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Impact of Effectors on the Catalytic Activity of Galactonolactone Oxidase from Trypanosoma cruziстатья Исследовательская статья

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