激活域和协同激活器之间的直接相互作用的高通量亲和度测量
Nicole DelRosso1, Peter H Suzuki2, Daniel Griffith3,4
1Biophysics Program, Stanford University, Stanford, CA, USA.
bioRxiv : the preprint server for biology
|September 4, 2024
概括
研究人员在转录因子激活域和协同激活剂之间绘制了1500多种蛋白质相互作用. 这揭示了新的结合对,并解释了这些相互作用如何控制基因激活.
科学领域:
- 分子生物学分子生物学
- 遗传学 遗传学 是一个
- 生物化学 生物化学
背景情况:
- 转录因子使用激活域来招募共同激活体,这是基因调节的关键步骤.
- 这些相互作用往往是弱的和短暂的,使得它们很难通过实验来研究.
- 了解这些相互作用是解读基因激活机制的关键.
研究的目的:
- 开发一种高通量方法,用于测量激活域和协同激活器之间的结合亲缘关系.
- 创建人类激活域-协同激活器相互作用的第一个定量地图.
- 研究控制这些相互作用的生物物理性质及其对基因激活的影响.
主要方法:
- 开发了一个微流体平台 (STAMMPPING) 用于并行表达和净化激活域.
- 在204个人类激活域和8个协同激活剂之间量化结合亲缘关系 (Kds).
- 利用结合多价值的分析模型来预测联合激活剂结合和细胞活动.
主要成果:
- 产生了超过1500个定量结合亲和关系,揭示了334个新的相互作用对.
- 确定共激活剂P300结合超过100个激活域.
- 证明了体外亲缘关系可以准确预测体内转录激活与超敏感反应.
结论:
- STAMMPPING平台使弱蛋白与蛋白相互作用的高通量测量成为可能.
- 生成的交互地图为理解转录调节提供了一个基础资源.
- 激活域的独特生物物理特性有助于特定的协同激活器招募和基因激活调整.
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