将细菌微分区外蛋白与遗传编码的冷凝物交接起来.
Michele Costantino1, Eric J Young2,3, Abesh Banerjee1
1School of Molecular Sciences, Arizona State University, Tempe, Arizona, USA.
细菌微分区 (BMC-H) 蛋白覆盖合成蛋白质凝聚物,形成稳定可控制的分区. 这一进步为设计具有可调节性质的合成细胞和器官提供了一种新方法.
科学领域:
- 生物化学 生物化学
- 合成生物学 合成生物学
- 生物物理学的生物物理.
背景情况:
- 液-液相分离 (LLPS) 驱动细胞凝结物的形成,这对合成细胞发育至关重要.
- 众所周知,细菌微分区 (BMC) 蛋白形成了强大的蛋白质结构.
研究的目的:
- 为了研究BMC-H蛋白在LLPS驱动的蛋白质凝聚物的组装.
- 设计BMC-H变种用于凝结物的表面涂层.
- 评估这些涂层冷凝剂的稳定性和功能性质,用于合成生物学应用.
主要方法:
- 利用由协同RGG-RGG域形成的工程蛋白质凝聚物.
- 工程 BMC-H 变种与 RGG 截断合并,以促进表面组装.
- 研究了pH和蛋白质度对涂层形成的影响.
- 评估了凝聚液凝聚,稳定性和选择性蛋白质结合 (例如TEV蛋白酶).
主要成果:
- 工程 BMC-H 变种,特别是 BMC-H-T2,在 RGG-RGG 水滴上形成了稳定的表面涂层.
- 这些涂层防止了滴滴凝聚,并且稳定,不会与稀释阶段交换.
- 涂层滴滴选择性地隔离了折叠的蛋白质,如TEV蛋白酶.
- TEV蛋白酶降解了RGG-RGG核心,但没有降解BMC-H涂层,证明了涂层的完整性.
结论:
- 贝蛋白质涂层的BMC蛋白质凝聚物是完全可编码的,可以精确控制LLPS区间.
- 该系统为构建功能合成器官和细胞提供了一个新的平台.
- 形成稳定,选择性涂层的能力为先进的生物分子工程开辟了道路.
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