A Structural and Functional Comparison Between Infectious and Non-Infectious Autocatalytic Recombinant PrP Conformers

PLoS Pathogens

Justin R. Barone¹  Michael B. Miller²  Geoffrey P. Noble²  Daniel J. Walsh²  Koren A. Nishina²  Surachai Supattapone²  Sheng Li³  Daphne W. Wang³ 

[1] Department of Biological Systems Engineering, Virginia Tech, Blacksburg, Virginia, United States of America;Departments of Biochemistry and Medicine, Geisel School of Medicine at Dartmouth, Hanover, New Hampshire, United States of America;Medicine and Biomedical Sciences Graduate Program, University of California at San Diego, La Jolla, California, United States of America

关键词: Cofactors (biochemistry);    Recombinant proteins;    Prions;    Prion diseases;    Deuterium;    Recombination reactions;    Sonication;    Post-translational modification;   

DOI  :  10.1371/journal.ppat.1005017

学科分类：生物科学（综合）

来源: Public Library of Science

【 摘 要 】

Infectious prions contain a self-propagating, misfolded conformer of the prion protein termed PrPSc. A critical prediction of the protein-only hypothesis is that autocatalytic PrPSc molecules should be infectious. However, some autocatalytic recombinant PrPSc 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 >105-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 PrPC substrates containing a glycosylphosphatidylinositol (GPI) anchor, including native PrPC. Taken together, these results indicate that having a conformation that can be specifically adopted by post-translationally modified PrPC molecules is an essential determinant of biological infectivity for recombinant prions, and suggest that this ability is associated with discrete features of PrPSc structure.

【 授权许可】

CC BY