Skip to main content
. 2021 Oct 25;297(5):101344. doi: 10.1016/j.jbc.2021.101344

Figure 5.

Figure 5

The influence of NPR on recPrP–POPG interaction.A, salt competition analysis. WT-recPrP and POPG were separately incubated with 650, 750, or 850 mM KCl before being mixed and analyzed in the iodixanol density gradient centrifugation. The gradient was collected from top to bottom for a total of 12 fractions. Control-N: WT-recPrP without POPG incubation. Control-P: WT-recPrP with POPG incubation. B, densitometric analysis of the PrP bands in (A). The binding of control-P was set as 100%. C, to compare the POPG binding between WT and NPR mutants, 750 mM KCl competed binding of WT recPrP to POPG (fourth panel in A) was compared directly with that of Met4-1 and ΔN6 recPrPs, which went through the same procedure. D, densitometric analysis of Western blots as shown in C (n = 4). E, preformed WT-POPG, Met4-1-POPG, or ΔN6-POPG complex was incubated with 1.5 M KCl and 10 mM NaOH prior to the gradient analysis. F, densitometric analysis of Western blots as shown in E (n = 4). In all panels, PrP was detected by Western blotting with the 3F10 anti-PrP antibody. Statistical significance was determined by one-way ANOVA followed by Tukey's multiple comparison test. ∗ represents p < 0.05; ∗∗∗ represents p < 0.01; ns represents no statistically significant difference. Error bars indicate standard deviations. Every point in D and F was from an independent experiment. NPR, N-terminal polybasic region of PrP; POPG, 1-palmitoyl-2-oleoylphosphatidylglycerol; recPrP, recombinant PrP.