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

IF 3.3 3区生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimie Pub Date : 2023-12-20 DOI:10.1016/j.biochi.2023.12.005

Valeriya V. Mikhaylova, Tatiana B. Eronina

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Abstract

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.

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拥挤条件下渗透溶质对肌肉糖原磷酸化酶b特性的影响

研究酶在渗透溶质存在的拥挤条件下的活性和稳定性之间的关系对于了解活细胞的功能非常重要。使用动态光散射、差示扫描量热仪、圆二色光谱、荧光测定法和酶活性测定法，研究了在 PEG 2000 和 PEG 20000 造成的拥挤条件下，渗透溶质（曲哈糖和甜菜碱）对肌肉糖原磷酸化酶 b（Phb）的二级和三级结构及活性的影响。25 °C 时，PEG 增加了 Phb 活性，但 PEG 20000 增加的程度更大。与 PEG 2000 相比，PEG 20000 明显破坏了其三级和二级结构的稳定性。海藻糖消除了 PEG 对 Phb 的影响，而甜菜碱则显著降低了 PEG 20000 的激活作用，但不影响 PEG 2000 的作用。在 48 °C 的热应激条件下，渗透溶质在拥挤条件下的保护作用比室温下更明显，在渗透溶质存在的条件下，Phb 的活性高于稀释溶液。这些结果为我们深入了解渗透溶质在拥挤条件下影响 Phb 的结构和活性的复杂机制提供了重要依据。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Biochimie 生物-生化与分子生物学

CiteScore

7.20

自引率

2.60%

发文量

219

审稿时长

40 days

期刊介绍： Biochimie publishes original research articles, short communications, review articles, graphical reviews, mini-reviews, and hypotheses in the broad areas of biology, including biochemistry, enzymology, molecular and cell biology, metabolic regulation, genetics, immunology, microbiology, structural biology, genomics, proteomics, and molecular mechanisms of disease. Biochimie publishes exclusively in English. Articles are subject to peer review, and must satisfy the requirements of originality, high scientific integrity and general interest to a broad range of readers. Submissions that are judged to be of sound scientific and technical quality but do not fully satisfy the requirements for publication in Biochimie may benefit from a transfer service to a more suitable journal within the same subject area.