The Structural Architecture of an Infectious Mammalian Prion Using Electron Cryomicroscopy
Vázquez-Fernández, Ester (University of Alberta. Centre for Prions and Protein Folding Diseases)
Vos, Matthijn R. (FEI Company)
Afanasyev, Pavel (Maastricht University. The Maastricht Multimodal Molecular Imaging Institute)
Cebey, Lino (University of Alberta. Centre for Prions and Protein Folding Diseases)
Sevillano, Alejandro M. (Universidade de Santiago de Compostela)
Vidal Barba, Enric (Institut de Recerca i Tecnologia Agroalimentàries. Centre de Recerca en Sanitat Animal)
Rosa, Isaac (Universidade de Santiago de Compostela)
Renault, Ludovic (University of Alberta. Department of Biochemistry)
Ramos, Adriana (Universidade de Santiago de Compostela)
Peters, Peter J. (Maastricht University. The Maastricht Multimodal Molecular Imaging Institute)
Fernández, José Jesús (Centro Nacional de Biotecnologia - CSIC)
van Heel, Marin (Imperial College London. Faculty of Natural Science)
Young, Howard S. (University of Alberta. Department of Biochemistry)
Requena, Jesús R. (Universidade de Santiago de Compostela)
Wille, Holger (University of Alberta. Centre for Prions and Protein Folding Diseases)

Date:	2016
Abstract:	The structure of the infectious prion protein (PrP Sc), which is responsible for Creutzfeldt-Jakob disease in humans and bovine spongiform encephalopathy, has escaped all attempts at elucidation due to its insolubility and propensity to aggregate. PrP Sc replicates by converting the non-infectious, cellular prion protein (PrP C) into the misfolded, infectious conformer through an unknown mechanism. PrP Sc and its N-terminally truncated variant, PrP 27-30, aggregate into amorphous aggregates, 2D crystals, and amyloid fibrils. The structure of these infectious conformers is essential to understanding prion replication and the development of structure-based therapeutic interventions. Here we used the repetitive organization inherent to GPI-anchorless PrP 27-30 amyloid fibrils to analyze their structure via electron cryomicroscopy. Fourier-transform analyses of averaged fibril segments indicate a repeating unit of 19. 1 Å. 3D reconstructions of these fibrils revealed two distinct protofilaments, and, together with a molecular volume of 18,990 Å 3, predicted the height of each PrP 27-30 molecule as ~17. 7 Å. Together, the data indicate a four-rung β-solenoid structure as a key feature for the architecture of infectious mammalian prions. Furthermore, they allow to formulate a molecular mechanism for the replication of prions. Knowledge of the prion structure will provide important insights into the self-propagation mechanisms of protein misfolding. The structure of the infectious prion (PrP Sc), which is responsible for Creutzfeldt-Jakob disease in humans and bovine spongiform encephalopathy, has escaped all attempts at elucidation due to its propensity to aggregate. Here, we use the repetitive organization inherent in amyloid fibrils to analyze the structure of GPI-anchorless PrP 27-30 via electron cryomicroscopy. Fourier-transform analysis of averaged fibril segments indicates a repeating unit of 19. 1 Å. In agreement with this observation, 3D reconstructions reveal that each fibril contains two distinct protofilaments and that the height of each PrP 27-30 molecule in these fibrils is ~17. 7 Å. Together the data indicate a four-rung β-solenoid structure as a key feature for the architecture of infectious mammalian prions. The data conflict with all previous models for the structure of PrP Sc and allow the formulation of a molecular mechanism for the replication of prions.
Grants:	Ministerio de Educación y Ciencia BFU2006-04588 Ministerio de Economía y Competitividad BFU2013-48436-C2-1-P Ministerio de Economía y Competitividad TIN2012-37483-C03-02
Rights:	Aquest document està subjecte a una llicència d'ús Creative Commons. Es permet la reproducció total o parcial, la distribució, la comunicació pública de l'obra i la creació d'obres derivades, fins i tot amb finalitats comercials, sempre i quan es reconegui l'autoria de l'obra original.
Language:	Anglès
Document:	Article ; recerca ; Versió publicada
Published in:	PLOS pathogens, Vol. 12 (september 2016) , ISSN 1553-7374

DOI: 10.1371/journal.ppat.1005835
PMID: 27606840

21 p, 24.0 MB

The record appears in these collections:
Research literature > UAB research groups literature > Research Centres and Groups (research output) > Health sciences and biosciences > Centre de Recerca en Sanitat Animal (CReSA-IRTA)
Articles > Research articles
Articles > Published articles