Fig. 3
Representative examples of distinct electrophoretic mobility patterns observed following proteinase K digestion of spontaneously misfolded recombinant PrP in PMSA. Along multiple PMSA experiments, the spontaneous formation of rec-PrPres with distinguishable electrophoretic mobility patterns was noticed after electrophoresis and total protein staining. These patterns were consistently obtained with varying frequencies. Thorough a comprehensive analysis of the PMSA products generated, we identified four distinct potentially different conformers. After proteinase K (PK) digestion, each conformer exhibited characteristic proteolytic fragments. The gel displays the potential recombinant strains Ust01 and Ust02, which both showed a 16 kDa band, two approximately 10 kDa bands with differing relative intensities (with Ust01 bands being similar and Ust02 bands predominantly displaying the higher band), and fragments of approximately 5 kDa. Additionally, the Ust06 strain exhibited the 16 kDa, 10 kDa, and 5 kDa fragments, along with a mild band over 10 kDa. Lastly, the Ust09 strain displayed an intense 16 kDa fragment and a ladder-like pattern below. rec-PrP: Undigested recombinant PrP. MW: Molecular weight marker