Direct observation of heterogeneous formation of amyloid spherulites in real-time by super-resolution microscopy.

ABSTRACT: Protein misfolding in the form of fibrils or spherulites is involved in a spectrum of pathological abnormalities. Our current understanding of protein aggregation mechanisms has primarily relied on the use of spectrometric methods to determine the average growth rates and diffraction-limited microscopes with low temporal resolution to observe the large-scale morphologies of intermediates. We developed a REal-time kinetics via binding and Photobleaching LOcalization Microscopy (REPLOM) super-resolution method to directly observe and quantify the existence and abundance of diverse aggregate morphologies of human insulin, below the diffraction limit and extract their heterogeneous growth kinetics. Our results revealed that even the growth of microscopically identical aggregates, e.g., amyloid

SUBMITTER: Zhang M 

PROVIDER: S-EPMC9392779 | biostudies-literature | 2022 Aug

REPOSITORIES: biostudies-literature