Insights into the Mechanism of Tryptophan Fluorescence Quenching due to Synthetic Crowding Agents: A Combined Experimental and Computational Study.
Insights into the Mechanism of Tryptophan Fluorescence Quenching due to Synthetic Crowding Agents: A Combined Experimental and Computational Study.
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DOI：
10.1021/acsomega.3c06006
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发表时间：
2023-11-28
期刊：
影响因子：
4.1
通讯作者：
Hati, Sanchita
中科院分区：
文献类型：
--
作者：
Fossum, Carl J.;Johnson, Benjamin O. V.;Golde, Spencer T.;Kielman, Alexis J.;Finke, Brianna;Smith, Macey A.;Lowater, Harrison R.;Laatsch, Bethany F.;Bhattacharyya, Sudeep;Hati, Sanchita
固有色氨酸荧光光谱是研究分子拥挤和限制对蛋白质结构、动力学和功能影响的重要工具。合成的拥挤物如葡聚糖、聚蔗糖、聚乙二醇、聚乙烯吡咯烷酮和它们各自的单体用于模拟拥挤的细胞内环境。因此，这些合成拥挤与色氨酸的相互作用及其对荧光特性的后续影响对于理解这些拥挤产生的可能干扰至关重要。在本研究中，聚合物和单体拥挤色氨酸荧光的影响进行了评估，通过使用实验和计算方法。本研究结果表明，聚合物和单体的拥挤对色氨酸的荧光强度有影响，但淬灭的分子机制是不同的。使用Stern-Volmer图和温度变化的研究，常用的合成crowders的淬火机制的物理基础。发现游离色氨酸的淬灭涉及静态、动态和作用范围机制。同时，采用Kohn-Sham密度泛函理论的计算研究提供了对分子间相互作用和溶剂化作用的影响的更深入的了解，从而导致这些拥挤者的不同猝灭模式。总之，这项研究提供了一些常用的单体和聚合物合成人群的淬灭机制的新的物理见解。
Intrinsic tryptophan fluorescence spectroscopy is an important tool for examining the effects of molecular crowding and confinement on the structure, dynamics, and function of proteins. Synthetic crowders such as dextran, ficoll, polyethylene glycols, polyvinylpyrrolidone, and their respective monomers are used to mimic crowded intracellular environments. Interactions of these synthetic crowders with tryptophan and the subsequent impact on its fluorescence properties are therefore critically important for understanding the possible interference created by these crowders. In the present study, the effects of polymer and monomer crowders on tryptophan fluorescence were assessed by using experimental and computational approaches. The results of this study demonstrated that both polymer and monomer crowders have an impact on the tryptophan fluorescence intensity; however, the molecular mechanisms of quenching were different. Using Stern–Volmer plots and a temperature variation study, a physical basis for the quenching mechanism of commonly used synthetic crowders was established. The quenching of free tryptophan was found to involve static, dynamic, and sphere-of-action mechanisms. In parallel, computational studies employing Kohn–Sham density functional theory provided a deeper insight into the effects of intermolecular interactions and solvation, resulting in differing quenching modes for these crowders. Taken together, the study offers new physical insights into the quenching mechanisms of some commonly used monomer and polymer synthetic crowders.
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影响因子：
3
作者：
Schlamadinger DE;Kats DI;Kim JE
通讯作者：
Kim JE
影响因子：
2.9
作者：
Johnson, James M.;Sanford, Brianne L.;Strom, Alexander M.;Tadayon, Stephanie N.;Lehman, Brent P.;Zirbes, Arrianna M.;Bhattacharyya, Sudeep;Musier-Forsyth, Karin;Hati, Sanchita
通讯作者：
Hati, Sanchita
影响因子：
12.9
作者：
Hu QH;Williams MT;Shulgina I;Fossum CJ;Weeks KM;Adams LM;Reinhardt CR;Musier-Forsyth K;Hati S;Bhattacharyya S
通讯作者：
Bhattacharyya S
影响因子：
5.6
作者：
Ghisaidoobe AB;Chung SJ
通讯作者：
Chung SJ
影响因子：
2.9
作者：
Breydo, Leonid;Sales, Amanda E.;Uversky, Vladimir N.
通讯作者：
Uversky, Vladimir N.