• 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Journal of Molecular Biology >Mechanistic Inferences From Analysis of Measurements of Protein Phase Transitions in Live Cells
【24h】

Mechanistic Inferences From Analysis of Measurements of Protein Phase Transitions in Live Cells

机译：从活细胞中蛋白质相变测量分析的机械推理

获取原文
获取原文并翻译 | 示例
           
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

The combination of phase separation and disorder-to-order transitions can give rise to ordered, semicrystalline fibrillar assemblies that underlie prion phenomena namely, the non-Mendelian transfer of information across cells. Recently, a method known as Distributed Amphifluoric Forster Resonance Energy Transfer (DAmFRET) was developed to study the convolution of phase separation and disorder-to-order transitions in live cells. In this assay, a protein of interest is expressed to a broad range of concentrations and the acquisition of local density and order, measured by changes in FRET, is used to map phase transitions for different proteins. The high-throughput nature of this assay affords the promise of uncovering sequence-to-phase behavior relationships in live cells. Here, we report the development of a supervised method to obtain automated and accurate classifications of phase transitions quantified using the DAmFRET assay. Systems that we classify as undergoing two-state discontinuous transitions are consistent with prion-like behaviors, although the converse is not always true. We uncover well-established and surprising new sequence features that contribute to two-state phase behavior of prion-like domains. Additionally, our method enables quantitative, comparative assessments of sequence-specific driving forces for phase transitions in live cells. Finally, we demonstrate that a modest augmentation of DAmFRET measurements, specifically time-dependent protein expression profiles, can allow one to apply classical nucleation theory to extract sequence-specific lower bounds on the probability of nucleating ordered assemblies. Taken together, our approaches lead to a useful analysis pipeline that enables the extraction of mechanistic inferences regarding phase transitions in live cells. (C) 2021 The Author(s). Published by Elsevier Ltd.
机译：相分离和无序到有序转变的结合可以产生有序、半结晶的纤维组装体，它们是朊病毒现象的基础，即细胞间信息的非孟德尔传递。最近，人们发展了一种称为分布式双荧光Forster共振能量转移（DAmFRET）的方法来研究活细胞中相分离和无序-有序转变的卷积。在该分析中，感兴趣的蛋白质被表达到广泛的浓度范围内，通过FRET变化测量的局部密度和顺序的获取被用于绘制不同蛋白质的相变图。这种分析的高通量特性为揭示活细胞中的序列-相行为关系提供了希望。在此，我们报告了一种有监督的方法的发展，以获得使用DAmFRET分析量化的相变的自动化和准确分类。我们归类为经历两态不连续跃迁的系统与类朊病毒的行为是一致的，尽管反过来并不总是正确的。我们发现了成熟且令人惊讶的新序列特征，这些特征有助于朊病毒样结构域的两态相行为。此外，我们的方法能够定量、比较地评估活细胞中特定于序列的相变驱动力。最后，我们证明，适度增加DAmFRET测量值，特别是时间依赖性蛋白质表达谱，可以使我们应用经典成核理论来提取有序组装成核概率的序列特定下限。综上所述，我们的方法产生了一个有用的分析管道，能够提取有关活细胞相变的机械推论。（c）2021作者。爱思唯尔有限公司出版。

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号