Complex Photophysical Properties of K114 Make for a Versatile Fluorescent Probe for Amyloid Detection.
Protein aggregation is a hallmark of Alzheimer's disease (AD) and many other neurodegenerative disorders. Small organic fluorophores such as Congo Red preferentially bind to cross-β-sheet-rich deposits and have been used to label amyloid plaques and tau tangles in histological samples. However, distinguishing between different conformations of protein aggregates is not trivial. Using silkworm and spider silks (prototypical amyloids) and transgenic AD mouse (5XFAD) and human AD brain samples, we report how spectral confocal microscopy allowed for improved detection and differentiation of protein aggregates based on the unexpected photophysical behavior of the amyloid-specific dye K114. The pH and excitation power had pronounced effects on the emission spectrum and intensity of amyloid-bound K114 fluorescence. When bound to β-sheet-rich assemblies, the emission spectrum of K114 was governed by the local pH of the binding pockets much more than by the pH of the mounting medium, likely due to ionization of titratable phenols. Unexpectedly, exposure to high excitation power caused a permanent increase in fluorescence intensity and a spectral blue-shift. These light-induced fluorescence changes were dependent in a complex manner on laser power, exposure time, pH, and amyloid type examined. The above-mentioned phenomena were observed in silk fibers and Alzheimer brain sections from mouse and human, indicating that this may be a general characteristic of K114 when bound to tightly aggregated macromolecules. Potential mechanisms are discussed, likely involving photoinduced electron transfer. Our findings illustrate how the complex photophysical behavior of amyloid-bound K114 can be exploited for improved detection and differentiation of protein aggregates.
['Stepanchuk|Anastasiia A|AA|', 'Heyne|Belinda|B|0000-0003-1655-6719', 'Stys|Peter K|PK|0000-0003-0326-4194']
D000544:Alzheimer Disease D000682:Amyloid D016229:Amyloid beta-Peptides D000818:Animals D001921:Brain D005456:Fluorescent Dyes D051379:Mice D008822:Mice, Transgenic D058225:Plaque, Amyloid
fluorescent probes spectral confocal microscopy Alzheimer’s disease Amyloid dyes imaging pathology