Understanding the Formation of Apoferritin Amyloid Fibrils Jurado Palomares, Rocío Gálvez Rodríguez, Natividad We present the optimization of experimental conditions to yield long, rigid apoferritin protein amyloid fibrils, as well as the corresponding fibrillation pathway. Fibril growth kinetics was followed using atomic force microscopy (AFM), transmission electron microscopy (TEM), dynamic light scattering (DLS), circular dichroism (CD), fourier-transform infrared spectroscopy (FTIR), and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Among the morphologies identified, we show that the conditions result in small aggregates, as well as medium and long fibrils. Extended incubation times led to progressive unfolding and hydrolysis of the proteins into very short peptide fragments. AFM, SDS-PAGE, and CD support a universal common fibrillation mechanism in which hydrolyzed fragments play the central role. These collective results provide convincing evidence that protein unfolding and complete hydrolysis of the proteins into very short peptide sequences are essential for the formation of the final apoferritin amyloid-like fibrils. 2023-03-08T11:44:00Z 2023-03-08T11:44:00Z 2021-04-06 journal article Biomacromolecules 2021, 22, 2057−2066. [https://doi.org/10.1021/acs.biomac.1c00176] https://hdl.handle.net/10481/80470 10.1021/acs.biomac.1c00176 eng http://creativecommons.org/licenses/by/4.0/ open access Atribución 4.0 Internacional American Chemical Society