在拥挤的溶液中排除体积和弱相互作用调节内在无序尾巴的形状和RNA结合
Madison A Stringer1,2, Jasmine Cubuk1,2, J Jeremías Incicco1
1Department of Biochemistry and Molecular Biophysics, Washington University in St Louis, 660 Euclid Avenue, Saint Louis, Missouri 63110, United States.
The journal of physical chemistry. B
|June 22, 2023
概括
细胞拥挤会影响蛋白质结构和RNA结合. 高分子量PEG会崩无序的蛋白尾,而低分子量PEG会扩大它们,从而影响SARS-CoV-2核体蛋白相互作用.
科学领域:
- 生物物理学的生物物理.
- 结构生物学 结构生物学
- 分子生物学分子生物学
背景情况:
- 细胞环境挤满了宏分子,影响蛋白质的行为.
- 内在无序的蛋白质对细胞拥挤特别敏感.
- 在病毒RNA包装中,SARS-CoV-2核体蛋白质起着至关重要的作用.
研究的目的:
- 为了研究细胞拥挤对SARS-CoV-2核体蛋白质内在无序的N-终端尾部的影响.
- 分析拥挤如何影响核体蛋白和RNA之间的相互作用.
- 了解拥挤在调节核体蛋白质变体之间的差异中的作用.
主要方法:
- 使用聚乙烯糖醇 (PEG) 模拟细胞拥挤.
- 使用单分子弗斯特共振能量转移 (smFRET) 来研究蛋白质构造.
- 在不同拥挤条件和温度下测量蛋白质-RNA结合亲和力.
主要成果:
- 高分子量PEG诱导了失调尾部的崩,而低分子量PEG则导致扩张.
- 拥挤剂增加了蛋白质-RNA结合亲和力,但这种效应在PEG分子量方面是非单调的.
- 拥挤减少了SARS-CoV-2核囊蛋白自然发生变体之间的亲和力差异.
结论:
- 细胞拥挤通过吸引力和排斥力的平衡显著影响内在无序的蛋白质区域.
- 拥挤调节蛋白质-连接体相互作用,对病毒RNA结合有影响.
- 了解拥挤效应对于准确模拟体内蛋白质行为以及解释生物变异至关重要.
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