Three-step Förster resonance energy transfer on an amyloid fibril scaffold.

The present study provides evidence that the energy transfer chain consisting of the benzothiazole dye Thioflavin T as an input donor, a phosphonium dye TDV and a squaraine dye SQ4 as mediators, and one of the three squaraines SQ1/2/3 as an output acceptor displays an excellent amyloid-sensing ability when applied to differentiating between the amyloid and non-fibrillized states of insulin. The ensemble of fluorophores offers the advantages of a large effective Stokes shift (∼240 nm), well-resolved 3D fluorescence patterns and strong enhancement of the terminal fluorescence (up to two orders of magnitude). The occurrence of multistep energy transfer on an amyloid fibril scaffold opens new possibilities for the more sensitive detection of fibrillar protein assemblies and their applications in nanophotonics.