Recombinant PrPScshares structural features with brain-derived PrPSc: Insights from limited proteolysis

Alejandro M. Sevillano, Natalia Fernández-Borges, Neelam Younas, Fei Wang, Saioa R. Elezgarai, Susana Bravo, Ester Vázquez-Fernández, Isaac Rosa, Hasier Eraña, David Gil, Sonia Veiga, Enric Vidal, Melissa L. Erickson-Beltran, Esteban Guitián, Christopher J. Silva, Romolo Nonno, Jiyan Ma, Joaquín Castilla, Jesús R. Requena

    Research output: Contribution to journalArticleResearchpeer-review

    11 Citations (Scopus)

    Abstract

    © 2018 Public Library of Science. All Rights Reserved. Very solid evidence suggests that the core of full length PrPScis a 4-rung β-solenoid, and that individual PrPScsubunits stack to form amyloid fibers. We recently used limited proteolysis to map the β-strands and connecting loops that make up the PrPScsolenoid. Using high resolution SDS-PAGE followed by epitope analysis, and mass spectrometry, we identified positions ~116/118, 133–134, 141, 152–153, 162, 169 and 179 (murine numbering) as Proteinase K (PK) cleavage sites in PrPSc. Such sites likely define loops and/or borders of β-strands, helping us to predict the threading of the β-solenoid. We have now extended this approach to recombinant PrPSc(recPrPSc). The term recPrPScrefers to bona fide recombinant prions prepared by PMCA, exhibiting infectivity with attack rates of ~100%. Limited proteolysis of mouse and bank vole recPrPScspecies yielded N-terminally truncated PK-resistant fragments similar to those seen in brain-derived PrPSc, albeit with varying relative yields. Along with these fragments, doubly N- and C-terminally truncated fragments, in particular ~89/97-152, were detected in some recPrPScpreparations; similar fragments are characteristic of atypical strains of brain-derived PrPSc. Our results suggest a shared architecture of recPrPScand brain PrPScprions. The observed differences, in particular the distinct yields of specific PK-resistant fragments, are likely due to differences in threading which result in the specific biochemical characteristics of recPrPSc. Furthermore, recombinant PrPScoffers exciting opportunities for structural studies unachievable with brain-derived PrPSc.
    Original languageEnglish
    Article numbere1006797
    JournalPLoS Pathogens
    Volume14
    Issue number1
    DOIs
    Publication statusPublished - 1 Jan 2018

    Fingerprint

    Dive into the research topics of 'Recombinant PrP<sup>Sc</sup>shares structural features with brain-derived PrP<sup>Sc</sup>: Insights from limited proteolysis'. Together they form a unique fingerprint.

    Cite this