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Structural and dynamical determinants of a β-sheet-enriched intermediate involved in amyloid fibrillar assembly of human prion protein
The conformational conversion of the cellular prion protein (PrP(C)) into a misfolded, aggregated and infectious scrapie isoform is associated with prion disease pathology and neurodegeneration. Despite the significant number of experimental and theoretical studies the molecular mechanism regulating...
Autores principales: | , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
The Royal Society of Chemistry
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9473526/ https://www.ncbi.nlm.nih.gov/pubmed/36277622 http://dx.doi.org/10.1039/d2sc00345g |
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author | Russo, Luigi Salzano, Giulia Corvino, Andrea Bistaffa, Edoardo Moda, Fabio Celauro, Luigi D'Abrosca, Gianluca Isernia, Carla Milardi, Danilo Giachin, Gabriele Malgieri, Gaetano Legname, Giuseppe Fattorusso, Roberto |
author_facet | Russo, Luigi Salzano, Giulia Corvino, Andrea Bistaffa, Edoardo Moda, Fabio Celauro, Luigi D'Abrosca, Gianluca Isernia, Carla Milardi, Danilo Giachin, Gabriele Malgieri, Gaetano Legname, Giuseppe Fattorusso, Roberto |
author_sort | Russo, Luigi |
collection | PubMed |
description | The conformational conversion of the cellular prion protein (PrP(C)) into a misfolded, aggregated and infectious scrapie isoform is associated with prion disease pathology and neurodegeneration. Despite the significant number of experimental and theoretical studies the molecular mechanism regulating this structural transition is still poorly understood. Here, via Nuclear Magnetic Resonance (NMR) methodologies we investigate at the atomic level the mechanism of the human HuPrP(90–231) thermal unfolding and characterize the conformational equilibrium between its native structure and a β-enriched intermediate state, named β-PrPI. By comparing the folding mechanisms of metal-free and Cu(2+)-bound HuPrP(23–231) and HuPrP(90–231) we show that the coupling between the N- and C-terminal domains, through transient electrostatic interactions, is the key molecular process in tuning long-range correlated μs–ms dynamics that in turn modulate the folding process. Moreover, via thioflavin T (ThT)-fluorescence fibrillization assays we show that β-PrPI is involved in the initial stages of PrP fibrillation, overall providing a clear molecular description of the initial phases of prion misfolding. Finally, we show by using Real-Time Quaking-Induced Conversion (RT-QuIC) that the β-PrPI acts as a seed for the formation of amyloid aggregates with a seeding activity comparable to that of human infectious prions. |
format | Online Article Text |
id | pubmed-9473526 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | The Royal Society of Chemistry |
record_format | MEDLINE/PubMed |
spelling | pubmed-94735262022-10-20 Structural and dynamical determinants of a β-sheet-enriched intermediate involved in amyloid fibrillar assembly of human prion protein Russo, Luigi Salzano, Giulia Corvino, Andrea Bistaffa, Edoardo Moda, Fabio Celauro, Luigi D'Abrosca, Gianluca Isernia, Carla Milardi, Danilo Giachin, Gabriele Malgieri, Gaetano Legname, Giuseppe Fattorusso, Roberto Chem Sci Chemistry The conformational conversion of the cellular prion protein (PrP(C)) into a misfolded, aggregated and infectious scrapie isoform is associated with prion disease pathology and neurodegeneration. Despite the significant number of experimental and theoretical studies the molecular mechanism regulating this structural transition is still poorly understood. Here, via Nuclear Magnetic Resonance (NMR) methodologies we investigate at the atomic level the mechanism of the human HuPrP(90–231) thermal unfolding and characterize the conformational equilibrium between its native structure and a β-enriched intermediate state, named β-PrPI. By comparing the folding mechanisms of metal-free and Cu(2+)-bound HuPrP(23–231) and HuPrP(90–231) we show that the coupling between the N- and C-terminal domains, through transient electrostatic interactions, is the key molecular process in tuning long-range correlated μs–ms dynamics that in turn modulate the folding process. Moreover, via thioflavin T (ThT)-fluorescence fibrillization assays we show that β-PrPI is involved in the initial stages of PrP fibrillation, overall providing a clear molecular description of the initial phases of prion misfolding. Finally, we show by using Real-Time Quaking-Induced Conversion (RT-QuIC) that the β-PrPI acts as a seed for the formation of amyloid aggregates with a seeding activity comparable to that of human infectious prions. The Royal Society of Chemistry 2022-08-05 /pmc/articles/PMC9473526/ /pubmed/36277622 http://dx.doi.org/10.1039/d2sc00345g Text en This journal is © The Royal Society of Chemistry https://creativecommons.org/licenses/by-nc/3.0/ |
spellingShingle | Chemistry Russo, Luigi Salzano, Giulia Corvino, Andrea Bistaffa, Edoardo Moda, Fabio Celauro, Luigi D'Abrosca, Gianluca Isernia, Carla Milardi, Danilo Giachin, Gabriele Malgieri, Gaetano Legname, Giuseppe Fattorusso, Roberto Structural and dynamical determinants of a β-sheet-enriched intermediate involved in amyloid fibrillar assembly of human prion protein |
title | Structural and dynamical determinants of a β-sheet-enriched intermediate involved in amyloid fibrillar assembly of human prion protein |
title_full | Structural and dynamical determinants of a β-sheet-enriched intermediate involved in amyloid fibrillar assembly of human prion protein |
title_fullStr | Structural and dynamical determinants of a β-sheet-enriched intermediate involved in amyloid fibrillar assembly of human prion protein |
title_full_unstemmed | Structural and dynamical determinants of a β-sheet-enriched intermediate involved in amyloid fibrillar assembly of human prion protein |
title_short | Structural and dynamical determinants of a β-sheet-enriched intermediate involved in amyloid fibrillar assembly of human prion protein |
title_sort | structural and dynamical determinants of a β-sheet-enriched intermediate involved in amyloid fibrillar assembly of human prion protein |
topic | Chemistry |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9473526/ https://www.ncbi.nlm.nih.gov/pubmed/36277622 http://dx.doi.org/10.1039/d2sc00345g |
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