Parameters that affect macromolecular self-assembly of prion protein

Amyloidogenesis of prion protein (PrP) is closely associated with the pathobiology of prion diseases. To understand details on formation of PrP amyloids, we investigated various conditions that influence the process in vitro, using full length and truncated recombinant PrP. Disrupted agitation and fluctuated temperature resulted in prolongation of lag phase during PrP amyloid formation. With the same conditions and material for the assay, fluorescence microplate readers of different manufacturers, which are assumed to have incongruent level of mechanical performance, demonstrated variations for the length of lag phase and the level of fluorescence detection. Presence of preformed amyloid seeds accelerated PrP amyloid formation. Similarly, recombinant proteins of different species affected effectual generation of amyloids. This process was also influenced by the concentrations and truncation of recombinant PrP. By investigating several conditions to perform PrP amyloid formation assay, our study addresses the factors that determine how much and how rapidly PrP amyloids are formed.