Sialylation of Prion Protein Controls the Rate of Prion Amplification, the Cross-Species Barrier, the Ratio of PrPSc Glycoform and Prion Infectivity
Figure 2
De-sialylation of PrPC increases the rate of PrPSc amplification in PMCAb.
A. 263K, HY, Drowsy or SSLOW scrapie brain materials were diluted 103–1010-fold into 10% NBH, dsNBH or mock treated NBH (NBH incubated with a buffer for enzymatic de-sialylation in the absence of sialidase) as indicated and subjected to a single PMCAb round. Undigested 10% NBH was used as reference. B. Analysis of PrPSc amplification fold. Scrapie brain materials were diluted 104-fold for 263K or 103-fold for SSLOW into 10% NBH or dsNBH as indicated and subjected to four serial PMCAb rounds. The material amplified in each round was diluted to a specified dilution fold into 10% NBH or dsNBH for the next PMCAb round as indicated. Unamplified seeds are shown as round 0. Prior to electrophoresis, samples were treated with PK. All blots were stained with 3F4 antibody.