单标记单分子荧光寿命实验中爆发内动力学的鉴定和定量。

IF 4.6 Q2 MATERIALS SCIENCE, BIOMATERIALS
ACS Applied Bio Materials Pub Date : 2022-09-14 Epub Date: 2022-08-17 DOI:10.1016/j.bpr.2022.100071
Paul David Harris, Eitan Lerner
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引用次数: 3

摘要

单分子光谱学彻底改变了分子生物物理学,并提供了探索生物分子内部结构部分如何在不同时间尺度上进行空间重组的方法。有几种单分子方法可以探测单个染料标记残留物附近的局部结构动力学,这些方法依赖于荧光寿命作为读数。然而,在微秒到毫秒的时间尺度上量化这种单分子单一染料荧光爆发动力学的分析框架尚未得到证实。在这里,我们提出了一个分析框架,用于识别和量化基于爆发寿命的动力学,例如单分子光异构化相关荧光增强中记录的构象动力学。在测试了模拟分析的能力之后,我们继续展示了未结合α-突触核蛋白单体在爆发毫秒内的局部结构动力学。本工作提供的分析框架为提取基于荧光寿命的单分子测量所探测的坐标的能量景观的全图铺平了道路。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

Identification and quantification of within-burst dynamics in singly labeled single-molecule fluorescence lifetime experiments.

Identification and quantification of within-burst dynamics in singly labeled single-molecule fluorescence lifetime experiments.

Identification and quantification of within-burst dynamics in singly labeled single-molecule fluorescence lifetime experiments.

Identification and quantification of within-burst dynamics in singly labeled single-molecule fluorescence lifetime experiments.

Single-molecule spectroscopy has revolutionized molecular biophysics and provided means to probe how structural moieties within biomolecules spatially reorganize at different timescales. There are several single-molecule methodologies that probe local structural dynamics in the vicinity of a single dye-labeled residue, which rely on fluorescence lifetimes as readout. Nevertheless, an analytical framework to quantify dynamics in such single-molecule single dye fluorescence bursts, at timescales of microseconds to milliseconds, has not yet been demonstrated. Here, we suggest an analytical framework for identifying and quantifying within-burst lifetime-based dynamics, such as conformational dynamics recorded in single-molecule photo-isomerization-related fluorescence enhancement. After testing the capabilities of the analysis on simulations, we proceed to exhibit within-burst millisecond local structural dynamics in the unbound α-synuclein monomer. The analytical framework provided in this work paves the way for extracting a full picture of the energy landscape for the coordinate probed by fluorescence lifetime-based single-molecule measurements.

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来源期刊
ACS Applied Bio Materials
ACS Applied Bio Materials Chemistry-Chemistry (all)
CiteScore
9.40
自引率
2.10%
发文量
464
期刊介绍: ACS Applied Bio Materials is an interdisciplinary journal publishing original research covering all aspects of biomaterials and biointerfaces including and beyond the traditional biosensing, biomedical and therapeutic applications. The journal is devoted to reports of new and original experimental and theoretical research of an applied nature that integrates knowledge in the areas of materials, engineering, physics, bioscience, and chemistry into important bio applications. The journal is specifically interested in work that addresses the relationship between structure and function and assesses the stability and degradation of materials under relevant environmental and biological conditions.
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