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Ruocheng Gu^1,2, Yunuo Lin^1,2, Rouyu Di^1,2

¹Department of Microbial Physiological & Metabolic Engineering, State Key Laboratory of Microbial Diversity and Innovative Utilization, Institute of Microbiology, Chinese Academy of Sciences, Beijing, 100101, China.

Advanced science (Weinheim, Baden-Wurttemberg, Germany)

|April 7, 2025

概括

工程整体可以实现高效的无痕蛋白质裂变,这是蛋白质结合的一个重大挑战. 该研究确定了一个关键的THN动机,对这种增强的C端裂解活动至关重要.

科学领域:

生物化学生物化学
分子生物学分子生物学
蛋白质工程是指蛋白质工程.

背景情况:

无痕蛋白质裂变对于蛋白应用至关重要,但由于当前蛋白活性和乱交的局限性,仍然具有挑战性.
现有的整因通常缺乏所需的效率或与各种N端氨基酸的兼容性,以实现无蛋白质结合.

研究的目的:

为了设计gp41-1中蛋白,使其具有高效且无痕迹的蛋白质裂变.
阐明工程智能中增强C端切割活动的结构和机制基础.

主要方法:

在gp41-1中使用的蛋白质工程.
评估裂变效率和与第一个氨基酸的兼容性.
使用分子动力学 (MD) 模拟来分析结构机制.
研究THN基因和特定氨基酸残留物的作用 (Thr123,His124,Asp107,Asn125).

主要成果:

工程设计的gp41-1整体表现出迄今为止报告的最高无痕切割效率.
在3类整体中保存的THN图案被确定为高效无痕C端切割的关键.
在THN基因内,一种特定的键 (Thr123-His124) 对于稳定基因和促进C端裂变至关重要.
突变Thr123显著破坏了THN动机,导致裂变活性丧失,实验数据和MD模拟证实了这一点.

结论:

THN图案是无痕切割效率的关键决定因素,特别是在C端.
了解THN图案的作用为设计具有优越裂解能力的新型内提供了宝贵的见解.
这项研究通过实现更有效的蛋白质结合策略,推进了蛋白质工程和生物技术中的整体的应用.

关键词:

这是一个THN图案.gp41‐1‐1 的时间.高分离活动的高分离活动.没有痕迹的裂纹.

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