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从折叠的折叠:探索单域蛋白质的拓异型

Zhiyu Qu^1,2, Lianjie Xu^1,2, Fengyi Jiang^1,2

¹Beijing National Laboratory for Molecular Sciences, Department of Polymer Science and Engineering, College of Chemistry and Molecular Engineering, Peking University, Beijing 100871, People's Republic of China.

Proceedings of the National Academy of Sciences of the United States of America

|October 15, 2024

概括

科学家们从单个蛋白质折叠中创建了七种蛋白质拓变异,包括节点和链接. 这些多样化的蛋白质结构共享一个核心,但表现出不同的折叠动态,扩大了我们对蛋白质结构的理解,超出了线性链.

科学领域:

蛋白质工程和结构生物学
生物物理学和分子动力学
合成生物学和蛋白质设计

背景情况:

探索超越线性排列的蛋白质拓对于理解蛋白质结构-功能关系至关重要.
由于其特定和快速结合能力,SpyTag-SpyCatcher系统为蛋白质工程提供了一个多功能平台.

研究的目的:

为了生成和表征单个蛋白质折叠的多样化的拓异型.
研究拓学对蛋白质结构,稳定性和动态的影响.
为研究超越线性链的蛋白质折叠景观建立一个模型系统.

主要方法:

通过重新连接二次结构元素和突变反应残留物来进行蛋白质工程.
使用蛋白质溶解消化,光相关谱学 (FCS) 和大小排除色谱学 (SEC) 的拓性表征.
通过差异扫描热度计 (DSC),NMR光谱 (HSQC-NMR) 和循环二元化 (CD) 光谱进行生物物理特征.
分子动力学 (MD) 模拟来分析展开的路径.

主要成果:

从一个蛋白质折叠前体成功创建了七种不同的拓异构形态 (线性,循环,结,拉索,伪托托克桑,) .
通过实验方法确认了拓,并证明了细胞表达.
所有异构体都具有稳定的疏水性核心,但表现出不同的折叠和展开动态.
实验和模拟数据揭示了蛋白质动态的拓依赖差异.

结论:

蛋白质折叠高度适应拓变异,使得从单个前体中创建多样化的结构.
拓学显著影响蛋白质折叠动态,同时保持核心结构完整性.
这项工作为蛋白质折叠景观提供了基本的见解,并为设计新型蛋白质架构打开了道路.

关键词:

模拟MD的模拟方法蛋白质设计蛋白质设计蛋白质的异构形式.蛋白质拓学的蛋白质拓学

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