PrPOligomersKineticData.zip

The mechanism of monomer transfer between two structurally distinct PrP oligomers, PLoS One 2017.

Full-length Ovine PrP 23-234 (Ala-136, Arg-154, Gln-171 variant) were produced in Escherichia coli and purified. The O1 oligomers were generated by incubating of OvPrP at 80mM at 55° C for 6 hours and purified by size exclusion chromatography (SEC). SEC experiments have been performed using TSK 4000 SW column equilibrated with Sodium citrate 20mM pH 4.0 coupled to an AKTA Purifier 100 (GE-HealthCare). The size distribution of O1 assemblies was estimated by coupling to multi-wavelength static light scattering with size exclusion chromatography using a TSK 4000SW. The resulting data were transformed to size distribution using a custom MATLAB program. The depolymerisation of O1 assemblies was followed by static light scattering (SLS) by incubating O1 assemblies at 50° C.