Xanthone-based solvatochromic fluorophores for quantifying micropolarity of protein aggregates.
Xanthone-based solvatochromic fluorophores for quantifying micropolarity of protein aggregates.
复制标题
DOI：
10.1039/d2sc05004h
复制
发表时间：
2022-11-02
期刊：
影响因子：
8.4
通讯作者：
中科院分区：
文献类型：
--
作者：
正确的三维结构对于维持蛋白质的功能以及避免错误折叠带来的病理后果至关重要。蛋白质的错误折叠和聚集都与神经退行性疾病有关。因此，人们开发了多种对极性和黏度都有响应的荧光工具来检测蛋白质聚集。然而，仅对极性有响应的高灵敏度荧光团的合理设计一直难以实现。在这项工作中，我们证明了具有（d - p） - π*共轭的吸电子杂原子能够稳定最低未占分子轨道（LUMO）能级并促进红移。在计算分析的指导下，我们设计了一系列新颖的基于呫吨酮的溶剂化变色荧光团，这些荧光团很少被系统研究。所得的探针对极性表现出极高的灵敏度，但对黏度不敏感。作为概念验证，我们合成了用于活细胞共聚焦成像的蛋白质靶向探针，旨在量化错误折叠和聚集蛋白质的极性。有趣的是，我们的结果以一种我们未曾预料到的方式揭示了蛋白质聚集的多个层面。首先，在折叠球状蛋白质的错误折叠和聚集过程中，极性降低的微环境得到了验证。其次，AgHalo的颗粒状聚集体比以Htt - polyQ为代表的折叠球状蛋白质形成的聚集体所呈现的环境极性更低。第三，我们的研究表明，因不同类型的应激源而形成的颗粒状蛋白质聚集体表现出显著的极性差异。这些结果表明，仅对极性有响应的溶剂化变色荧光团代表了一类新的指示剂，可广泛用于检测活细胞中的蛋白质聚集，从而为阐明蛋白质聚集的细胞机制以及处理细胞内聚集体的治疗方法铺平了道路。
通过单原子取代方法，我们开发了一系列仅对极性有响应的溶剂化变色荧光团。通过对错误折叠和聚集蛋白质的极性进行量化，证明了这些荧光团的实用性。
Proper three-dimensional structures are essential for maintaining the functionality of proteins and for avoiding pathological consequences of improper folding. Misfolding and aggregation of proteins have been both associated with neurodegenerative disease. Therefore, a variety of fluorogenic tools that respond to both polarity and viscosity have been developed to detect protein aggregation. However, the rational design of highly sensitive fluorophores that respond solely to polarity has remained elusive. In this work, we demonstrate that electron-withdrawing heteroatoms with (d–p)–π* conjugation can stabilize lowest unoccupied molecular orbital (LUMO) energy levels and promote bathochromic shifts. Guided by computational analyses, we have devised a novel series of xanthone-based solvatochromic fluorophores that have rarely been systematically studied. The resulting probes exhibit superior sensitivity to polarity but are insensitive to viscosity. As proof of concept, we have synthesized protein targeting probes for live-cell confocal imaging intended to quantify the polarity of misfolded and aggregated proteins. Interestingly, our results reveal several layers of protein aggregates in a way that we had not anticipated. First, microenvironments with reduced polarity were validated in the misfolding and aggregation of folded globular proteins. Second, granular aggregates of AgHalo displayed a less polar environment than aggregates formed by folded globular protein represented by Htt–polyQ. Third, our studies reveal that granular protein aggregates formed in response to different types of stressors exhibit significant polarity differences. These results show that the solvatochromic fluorophores solely responsive to polarity represent a new class of indicators that can be widely used for detecting protein aggregation in live cells, thus paving the way for elucidating cellular mechanisms of protein aggregation as well as therapeutic approaches to managing intracellular aggregates. Using a single atom substitution approach, we have developed a series of solvatochromic fluorophores that respond solely to polarity. The utility of these fluorophores is demonstrated by quantifying the polarity of misfolded and aggregated proteins.
登录
查看更多内容
影响因子：
8.4
作者：
Dziuba D;Pospíšil P;Matyašovský J;Brynda J;Nachtigallová D;Rulíšek L;Pohl R;Hof M;Hocek M
通讯作者：
Hocek M
影响因子：
8.4
作者：
Er, Jun Cheng;Tang, Mui Kee;Chang, Young-Tae
通讯作者：
Chang, Young-Tae
影响因子：
3.8
作者：
Hassan, Syed Imran;Haque, Ashanul;Mushtaque, Md.
通讯作者：
Mushtaque, Md.
影响因子：
2.9
作者：
Liu, Yu;Miao, Kun;Zhang, Xin
通讯作者：
Zhang, Xin
影响因子：
15
作者：
Liu Y;Wolstenholme CH;Carter GC;Liu H;Hu H;Grainger LS;Miao K;Fares M;Hoelzel CA;Yennawar HP;Ning G;Du M;Bai L;Li X;Zhang X
通讯作者：
Zhang X