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三氟乙酸作为液-液相分离肽系统中致密相的分子探针。

Trifluoroacetic Acid as a Molecular Probe for the Dense Phase in Liquid-Liquid Phase-Separating Peptide Systems.

作者信息

Lim Jessica, Chin SzeYuet, Miserez Ali, Xue Kai, Pervushin Konstantin

机构信息

School of Biological Sciences, Nanyang Technological University, 60 Nanyang Drive, 637551 Singapore.

Centre of High Field NMR Spectroscopy and Imaging, Nanyang Technological University, 21 Nanyang Link, 637371 Singapore.

出版信息

Anal Chem. 2025 Jan 14;97(1):166-174. doi: 10.1021/acs.analchem.4c03444. Epub 2024 Dec 22.

DOI:10.1021/acs.analchem.4c03444
PMID:39710972
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11740181/
Abstract

Although trifluoroacetic acid (TFA) is not typically considered a Hofmeister reagent, it has been demonstrated to modulate biocoacervation. We show that TFA can be employed to probe specific interactions in coacervating bioinspired peptide phenylalanine (Phe) F-labeled at a single site, altering its liquid-liquid phase separation (LLPS) behavior. Solid-state nuclear magnetic resonance (NMR) spectroscopy revealed two dynamically distinct binding modes of TFA with Phe, resulting in a structured, dipolar-ordered complex and a more dynamic complex, highlighting the proximity between TFA and Phe. Quantum chemistry modeling of F chemical shift differences indicates that the structured complex is formed by the intercalation of one TFA molecule between two stacked Phe aromatic rings, possibly contributing to the stabilization of the condensed dense phase. Thus, we propose that TFA can be used as a convenient molecular probe in F NMR-based studies of the structure and dynamics of the dense phase in LLPS peptide systems.

摘要

尽管三氟乙酸（TFA）通常不被视为霍夫迈斯特试剂，但已证明它能调节生物凝聚作用。我们表明，TFA可用于探测在单个位点标记有生物启发肽苯丙氨酸（Phe）F的凝聚过程中的特定相互作用，改变其液-液相分离（LLPS）行为。固态核磁共振（NMR）光谱揭示了TFA与Phe的两种动态不同的结合模式，形成了一种结构化的、具有偶极有序的复合物和一种更具动态性的复合物，突出了TFA与Phe之间的接近程度。F化学位移差异的量子化学建模表明，结构化复合物是由一个TFA分子插入两个堆叠的Phe芳香环之间形成的，这可能有助于凝聚致密相的稳定。因此，我们提出TFA可作为一种方便的分子探针，用于基于F NMR的LLPS肽系统致密相结构和动力学研究。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a8d6/11740181/1593795291aa/ac4c03444_0005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a8d6/11740181/d7eeb8e72efc/ac4c03444_0001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a8d6/11740181/5da438dc7d4e/ac4c03444_0002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a8d6/11740181/6d5452605b7e/ac4c03444_0003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a8d6/11740181/cebe2ffdeb72/ac4c03444_0004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a8d6/11740181/1593795291aa/ac4c03444_0005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a8d6/11740181/d7eeb8e72efc/ac4c03444_0001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a8d6/11740181/5da438dc7d4e/ac4c03444_0002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a8d6/11740181/6d5452605b7e/ac4c03444_0003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a8d6/11740181/cebe2ffdeb72/ac4c03444_0004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a8d6/11740181/1593795291aa/ac4c03444_0005.jpg

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