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Updated: May 30, 2025

Measuring In Vitro ATPase Activity for Enzymatic Characterization

Measuring In Vitro ATPase Activity for Enzymatic Characterization

Published on: August 23, 2016

ATP 结合和抑制内在无序的蛋白相互作用

Shiyan Lin¹, Guorong Hu¹, Moxin Zhang¹

¹Zhejiang Province Key Laboratory of Quantum Technology and Device, School of Physics, Zhejiang University, Yuhangtang Road 866, Hangzhou 310058, China.

Langmuir : the ACS journal of surfaces and colloids

|January 31, 2025

概括

高度的腺三酸盐 (ATP) 抑制了内在无序蛋白质 (IDP) 中的液态-液态相分离 (LLPS). ATP 竞争性地与关键残留物结合,揭示了LLPS抑制的一般机制.

科学领域:

生物化学生化学
分子生物学分子生物学
生物物理学的生物物理.

背景情况:

内在无序的蛋白质 (IDPs) 驱动液体-液体相分离 (LLPS),对于细胞组织至关重要.
已知高生理度的腺三酸盐 (ATP) 抑制了IDP驱动的LLPS.
在LLPS上ATP的抑制作用背后的精确机制在很大程度上仍未被阐明.

研究的目的:

调查ATP抑制由IDP驱动的LLPS的分子机制.
通过计算模拟,阐明ATP与特定的IDP之间的相互作用动态.
为了确定竞争性结合是否是ATP介导的LLPS抑制的一般机制.

主要方法:

采用了全原子分子动力学 (MD) 模拟.
分析了ATP与两个代表性IDP的相互作用,即FUS PLD和hNRNP G RGG域.
评估了结合亲缘关系,关键相互作用残留物和动态过程.

主要成果:

ATP对两种研究的IDP都表现出强烈的结合亲和力,有效地抑制了它们的相互作用.
对于 hnRNP G RGG 域,ATP 具有竞争性地与氨酸残留物结合,这对于 IDP 相互作用至关重要.
在FUS PLD中,ATP也可以竞争性地与谷氨酸,氨酸和氨酸等残留物结合,尽管由于残留物丰富而存在温和的竞争. 在PLD表面的ATP扩散明显快于PLD接触区域的演变,增强了抑制.

结论:

对ATP与关键残留物的竞争性结合是抑制IDP相互作用和LLPS的主要机制.
ATP扩散和IDP相互作用动态之间的时间分离进一步增强了ATP对FUS PLD的抑制作用.
这种竞争性结合机制可能代表ATP诱导的LLPS抑制在高生理度的一般模式.

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