Structure of a pathogenic amyloid nucleus determined by rational genetic deconstruction of an intracellular nucleation barrier

Abstract

A long-standing goal of the study of amyloids has been to characterize the structural basis of the rate-determining nucleating event. However, the ephemeral nature of that event has made it inaccessible to classical biochemistry, structural biology, and computational approaches. Here, we addressed that limitation by using Distributed Amphifluoric FRET to measure the dependence of amyloid formation on concentration and conformational templates in living cells, whose volumes are sufficiently small to resolve the outcomes of independent nucleation events.