Quantifying the Role of Lysine in Prion Replication by Nano-LC Mass Spectrometry and Bioassay.

ABSTRACT: Prions propagate by a template driven process, inducing the normal cellular isoform (PrP^C) to adopt the prion (PrP^Sc) conformation. In PrP^C, the positions of lysines are highly conserved and strongly influence prion propagation. In this study, covalent modification was used to quantitate the role of lysines in the PrP^Sc template that drives prion replication. The ε-amino group of lysines in the PrP^Sc (hamster-adapted scrapie Sc237) template was acetylated by either acetic anhydride (Ac₂O) or the N-hydroxysuccinimide ester of acetic acid (Ac-NHS). The extent of lysine acetylation in PrP^Sc was quantitated by mass spectrometry or Western blot-based analysis. Identical samples were bioassayed to quantitate the loss of

SUBMITTER: Silva CJ 

PROVIDER: S-EPMC7542330 | biostudies-literature | 2020

REPOSITORIES: biostudies-literature