CAS Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, 457 Zhongshan Road, Dalian, 116023, China.
National Laboratory of Biomacromolecules, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, 15 Datun Road, Chaoyang District, Beijing, 100101, China.
Angew Chem Int Ed Engl. 2021 Dec 1;60(49):25865-25871. doi: 10.1002/anie.202107943. Epub 2021 Nov 2.
We report a crystallization-induced emission fluorophore to quantitatively interrogate the polarity of aggregated proteins. This solvatochromic probe, namely "AggRetina" probe, inherently binds to aggregated proteins and exhibits both a polarity-dependent fluorescence emission wavelength shift and a viscosity-dependent fluorescence intensity increase. Regulation of its polarity sensitivity was achieved by extending the conjugation length. Different proteins bear diverse polarity upon aggregation, leading to different resistance to proteolysis. Polarity primarily decreases during protein misfolding but viscosity mainly increases upon the formation of insoluble aggregates. We quantified the polarity of aggregated protein-of-interest in live cells via HaloTag bioorthogonal labeling, revealing polarity heterogeneity within cellular aggregates. The enriched micro-environment details inside misfolded and aggregated proteins may correlate to their bio-chemical properties and pathogenicity.
我们报告了一种晶致发光荧光团，可以定量研究聚集蛋白的极性。这种溶致变色探针，即“聚集视网膜”探针，固有地与聚集蛋白结合，并表现出对极性的依赖性荧光发射波长位移和对粘度的依赖性荧光强度增加。通过延长共轭长度来调节其极性敏感性。不同的蛋白质在聚集时具有不同的极性，导致对蛋白水解的不同抗性。极性主要在蛋白质错误折叠时降低，但在形成不溶性聚集体时粘度主要增加。我们通过 HaloTag 生物正交标记在活细胞中定量测定了感兴趣的聚集蛋白的极性，揭示了细胞聚集体内部的极性异质性。错误折叠和聚集蛋白内部丰富的微观环境细节可能与其生化特性和致病性相关。
