© 2015 Noble et al. Infectious prions contain a self-propagating, misfolded conformer of the prion protein termed PrP < sup > Sc < /sup > . A critical prediction of the protein-only hypothesis is that autocatalytic PrP < sup > Sc < /sup > molecules should be infectious. However, some autocatalytic recombinant PrP < sup > Sc < /sup > molecules have low or undetectable levels of specific infectivity in bioassays, and the essential determinants of recombinant prion infectivity remain obscure. To identify structural and functional features specifically associated with infectivity, we compared the properties of two autocatalytic recombinant PrP conformers derived from the same original template, which differ by > 10 < sup > 5 < /sup > -fold in specific infectivity for wild-type mice. Structurally, hydrogen/deuterium exchange mass spectrometry (DXMS) studies revealed that solvent accessibility profiles of infectious and non-infectious autocatalytic recombinant PrP conformers are remarkably similar throughout their protease-resistant cores, except for two domains encompassing residues 91-115 and 144-163. Raman spectroscopy and immunoprecipitation studies confirm that these domains adopt distinct conformations within infectious versus non-infectious autocatalytic recombinant PrP conformers. Functionally, in vitro prion propagation experiments show that the non-infectious conformer is unable to seed mouse PrP < sup > C < /sup > substrates containing a glycosylphosphatidylinositol (GPI) anchor, including native PrP < sup > C < /sup > . Taken together, these results indicate that having a conformation that can be specifically adopted by post-translationally modified PrP < sup > C < /sup > molecules is an essential determinant of biological infectivity for recombinant prions, and suggest that this ability is associated with discrete features of PrP < sup > Sc < /sup > structure.
Noble, G. P., Wang, D. W., Walsh, D. J., Supattapone, S., Miller, M. B., Nishina, K. A., … Supattapone, S. (2015). A Structural and Functional Comparison Between Infectious and Non-Infectious Autocatalytic Recombinant PrP Conformers. PLoS Pathogens, 11(6). https://doi.org/10.1371/journal.ppat.1005017