Modulation of Fluorescent Protein Chromophores To Detect Protein Aggregation with Turn-On Fluorescence

• DOI: 10.1021/jacs.8b02176
• Publication Date: 2018-06-08T18:27:43Z
• Authors: Yu Liu; Charles H. Wolste...; Gregory C. Carter; Hongbin Liu; Hang Hu; Leeann S. Grainger; Kun Miao; Matthew Fares; Conner A. Hoelzel; Hemant P. Yennawar; Gang Ning; Manyu Du; Lu Bai; Xiaosong Li; Xin Zhang
• Keywords: Chromophore; Bioorthogonal Chemistry; Bimolecular fluorescence complementation; Fluorescent protein

We present a fluorogenic method to visualize misfolding and aggregation of specific protein-of-interest in live cells using structurally modulated fluorescent protein chromophores. Combining photophysical analysis, X-ray crystallography, theoretical calculation, we show that fluorescence is triggered by inhibition twisted-intramolecular charge transfer these fluorophores the rigid microenvironment viscous solvent or aggregates. Bioorthogonal conjugation fluorophore Halo-tag fused protein-of-interests allows for detection both misfolded aggregated species cells. Unlike other methods, our capable detecting previously invisible soluble proteins. This work provides first application chromophores detect conformational collapse