Department of Biology, College of Arts and Sciences, University of Kentucky, 675 Rose St., Lexington, KY, 40506, USA.
Protein J. 2014 Jun;33(3):243-52. doi: 10.1007/s10930-014-9556-z.
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.
朊病毒蛋白（PrP）的淀粉样变性与朊病毒病的病理生物学密切相关。为了详细了解 PrP 淀粉样纤维的形成，我们使用全长和截断的重组 PrP 研究了影响体外过程的各种条件。搅拌中断和温度波动导致 PrP 淀粉样形成的迟滞期延长。在相同的条件和材料下，不同制造商的荧光微孔板读数器，其机械性能水平被认为不一致，表现出迟滞期长度和荧光检测水平的变化。预形成的淀粉样纤维种子的存在加速了 PrP 淀粉样纤维的形成。同样，不同物种的重组蛋白也会影响淀粉样纤维的有效生成。该过程还受到重组 PrP 的浓度和截断的影响。通过研究几种条件来进行 PrP 淀粉样形成测定，我们的研究解决了决定 PrP 淀粉样纤维形成量和速度的因素。
