Single-Molecule Reaction-Diffusion.
Single-Molecule Reaction-Diffusion.
复制标题
单分子反应扩散。
DOI：
10.1101/2023.09.05.556378
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发表时间：
2023
期刊：
影响因子：
--
通讯作者：
Pressé,Steve
中科院分区：
文献类型：
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作者：
Xu徐伟青,LanceWQ;Jazani,Sina;Kilic,Zeliha;Pressé,Steve
我们建议捕捉反应扩散的分子，从最快的可获得的时间尺度，即个人光子到达的分子的基础上。我们说明了我们的方法，内在的无序的人类蛋白质，连接器组蛋白H1.0以及其伴侣胸腺素原，因为这些扩散通过照明共聚焦点和相互作用，形成更大的三元复合物的毫秒时间尺度。最重要的是，单分子反应扩散，smRD，揭示了单分子的性质，没有捕获或以其他方式限制分子的表面。我们在贝叶斯范式内实现smRD，并将我们的方法称为贝叶斯smRD。贝叶斯-smRD是进一步免费的平均，批量，结果固有的长光子到达痕迹的荧光相关光谱分析。在从数以千计的光子到达连续的空间位置和离散的构象和非物理状态变化的学习，贝叶斯smRD估计动力学参数的分子的分子的基础上，两到三个数量级的数据比工具，如荧光相关光谱，从而也大大减少了样品的光损伤。
We propose to capture reaction-diffusion on a molecule-by-molecule basis from the fastest acquirable timescale, namely individual photon arrivals. We illustrate our method on intrinsically disordered human proteins, the linker histone H1.0 as well as its chaperone prothymosin, as these diffuse through an illuminated confocal spot and interact forming larger ternary complexes on millisecond timescales. Most importantly, single-molecule reaction-diffusion, smRD, reveals single molecule properties without trapping or otherwise confining molecules to surfaces. We achieve smRD within a Bayesian paradigm and term our method Bayes-smRD. Bayes-smRD is further free of the average, bulk, results inherent to the analysis of long photon arrival traces by fluorescence correlation spectroscopy. In learning from thousands of photon arrivals continuous spatial positions and discrete conformational and photophysical state changes, Bayes-smRD estimates kinetic parameters on a molecule-by-molecule basis with two to three orders of magnitude less data than tools such as fluorescence correlation spectroscopy thereby also dramatically reducing sample photodamage.