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Fibril paint Targets Amyloid Fibrils For Ubiquitination
This preprint describes Fibril Paint peptides that bind amyloid fibrils and recruit the E3 ubiquitin ligase CHIP, so that fibrils of Tau, huntingtin, α-synuclein and amyloid-β, including fibrils from patient brain, are tagged with ubiquitin as a first step towards their degradation. Flow Induced Dispersion Analysis (FIDA), on a Fida 1 instrument, was used to follow fluorescently labelled ubiquitin through the ubiquitination cascade, reading its hydrodynamic radius as it was passed to the E1 and E2 enzymes and finally attached to the fibrils. FIDA also showed that the ubiquitin stayed on the fibrils after denaturant treatment, consistent with covalent attachment. Binding of Fibril Paint to CHIP was measured by microscale thermophoresis, not by FIDA. It is an example of using FIDA to track an enzymatic cascade and the size of modified amyloid fibrils in solution.
