Effects of osmolytes under crowding conditions on the properties of muscle glycogen phosphorylase b.

Autor: Mikhaylova VV; Bach Institute of Biochemistry, Federal Research Centre 'Fundamentals of Biotechnology' of the Russian Academy of Sciences, Leninsky pr. 33, Moscow, 119071, Russia. Electronic address: mikhaylova.inbi@inbox.ru., Eronina TB; Bach Institute of Biochemistry, Federal Research Centre 'Fundamentals of Biotechnology' of the Russian Academy of Sciences, Leninsky pr. 33, Moscow, 119071, Russia.
Jazyk: angličtina
Zdroj: Biochimie [Biochimie] 2024 May; Vol. 220, pp. 48-57. Date of Electronic Publication: 2023 Dec 20.
DOI: 10.1016/j.biochi.2023.12.005
Abstrakt: The study of the relationship between the activity and stability of enzymes under crowding conditions in the presence of osmolytes is important for understanding the functioning of a living cell. The effect of osmolytes (trehalose and betaine) on the secondary and tertiary structure and activity of muscle glycogen phosphorylase b (Phb) under crowding conditions created by PEG 2000 and PEG 20000 was investigated using dynamic light scattering, differential scanning calorimetry, circular dichroism spectroscopy, fluorimetry and enzymatic activity assay. At 25 °C PEGs increased Phb activity, but PEG 20000 to a greater extent. Wherein, PEG 20000 significantly destabilized its tertiary and secondary structure, in contrast to PEG 2000. Trehalose removed the effects of PEGs on Phb, while betaine significantly reduced the activating effect of PEG 20000 without affecting the action of PEG 2000. Under heat stress at 48 °C, the protective effect of osmolytes under crowding conditions was more pronounced than at room temperature, and the Phb activity in the presence of osmolytes was higher in these conditions than in diluted solutions. These results provide important insights into the complex mechanism, by which osmolytes affect the structure and activity of Phb under crowding conditions.
Competing Interests: Declaration of competing interest The authors have declared that there are no conflicts of interest. The funders had no role in study design, data collection, analysis, interpretation, writing of the manuscript, or the decision to publish the results.
(Copyright © 2023 Elsevier B.V. and Société Française de Biochimie et Biologie Moléculaire (SFBBM). All rights reserved.)
Databáze: MEDLINE
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