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首页> 外文期刊>The journal of physical chemistry, A. Molecules, spectroscopy, kinetics, environment, & general theory >Quantitative single-molecule conformational distributions: A case study with poly-(L-proline)
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Quantitative single-molecule conformational distributions: A case study with poly-(L-proline)

机译：定量单分子构象分布：以聚（L-脯氨酸）为例

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摘要

Precise measurement of the potential of mean force is necessary for a fundamental understanding of the dynamics and chemical reactivity of a biological macromolecule. The unique advantage provided by the recently developed constant-information approach to analyzing time-dependent single-molecule fluorescence measurements was used with maximum entropy deconvolution to create a procedure for the accurate determination of molecular conformational distributions, and analytical expressions for the errors in these distributions were derived. This new method was applied to a derivatized poly(L-proline) series, P(n)CG(3)K-(biotin) (n = 8, 12, 15, 18, and 24), using a modular, server-based single-triolecule spectrometer that is capable of registering photon arrival times with a continuous-wave excitation Source. To account for potential influence from the microscopic environment, factors that were calibrated and corrected molecule by molecule include background, cross-talk, and detection efficiency. For each single poly(L-proline) molecule, sharply peaked Forster type resonance energy transfer (FRET) efficiency and distance distributions were recovered, indicating a static end-to-end distance on the time scale of measurement. The experimental distances were compared with models of varying rigidity. The results suggest that the 23 A persistence length wormlike chain model derived from experiments with high molecular weight poly(L-proline) is applicable to short chains as well.
机译：要准确了解生物大分子的动力学和化学反应性，必须精确测量平均力的潜力。最近开发的恒定信息方法分析时间相关的单分子荧光测量所提供的独特优势与最大熵解卷积一起使用，从而创建了精确确定分子构象分布的程序，并为这些分布中的误差提供了解析表达式被派生。这项新方法通过使用模块化的服务器端方法，应用于衍生化的聚（L-脯氨酸）系列P（n）CG（3）K-（生物素）（n = 8、12、15、18和24）。基于单分子的光谱仪，能够用连续波激发源记录光子到达时间。为了说明来自微观环境的潜在影响，逐个分子校准和校正的因素包括背景，串扰和检测效率。对于每个单个的聚（L-脯氨酸）分子，均恢复了峰峰值明显的Forster型共振能量转移（FRET）效率和距离分布，表明在测量时标上的静态端到端距离。将实验距离与不同刚度的模型进行比较。结果表明，从高分子量聚（L-脯氨酸）实验获得的23 A持久长度蠕虫状链模型也适用于短链。

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