Modulation of Fluorescent Protein Chromophores To Detect Protein Aggregation with Turn-On Fluorescence
| Autor: | Xiaosong Li, Gang Ning, Hemant P. Yennawar, Manyu Du, Gregory C. Carter, Yu Liu, Lu Bai, Hongbin Liu, Conner A. Hoelzel, Kun Miao, Matthew Fares, Xin Zhang, Leeann S. Grainger, Hang Hu, Charles H. Wolstenholme |
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| Jazyk: | angličtina |
| Rok vydání: | 2018 |
| Předmět: |
Protein Folding
Fluorophore 02 engineering and technology Protein aggregation 010402 general chemistry 01 natural sciences Biochemistry Catalysis Article Fluorescence Turn (biochemistry) chemistry.chemical_compound Colloid and Surface Chemistry Superoxide Dismutase-1 Humans Imidazolines Fluorescent Dyes Huntingtin Protein Microscopy Confocal HEK 293 cells General Chemistry Chromophore 021001 nanoscience & nanotechnology 0104 chemical sciences HEK293 Cells chemistry Microscopy Fluorescence Mutation Biophysics alpha-Synuclein Protein folding Bioorthogonal chemistry Protein Multimerization 0210 nano-technology |
| Popis: | We present a fluorogenic method to visualize misfolding and aggregation of a specific protein-of-interest in live cells using structurally modulated fluorescent protein chromophores. Combining photo-physical analysis, X-ray crystallography, and theoretical calculation, we show that fluorescence is triggered by inhibition of twisted-intramolecular charge transfer of these fluorophores in the rigid microenvironment of viscous solvent or protein aggregates. Bioorthogonal conjugation of the fluorophore to Halo-tag fused protein-of-interests allows for fluorogenic detection of both misfolded and aggregated species in live cells. Unlike other methods, our method is capable of detecting previously invisible misfolded soluble proteins. This work provides the first application of fluorescent protein chromophores to detect protein conformational collapse in live cells. |
| Databáze: | OpenAIRE |
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