Structural and dynamical determinants of a β-sheet-enriched intermediate involved in amyloid fibrillar assembly of human prion protein

Abstract

The conformational conversion of the cellular prion protein (PrPC) into 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 NMR methodologies we investigate at 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 Cu2+-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 first stages of PrP fibrillation, overall providing a clear molecular description of the first phases of prion misfolding. Finally, we show by using Real-Time Quaking-Induced Conversion (RT-QuIC) that the β-PrPI acts as seed for the formation of amyloid aggregates with a seeding activity comparable to that of human infectious prions.

Supplementary files

Article information

Article type
Edge Article
Submitted
18 Jan 2022
Accepted
04 Aug 2022
First published
05 Aug 2022
This article is Open Access

All publication charges for this article have been paid for by the Royal Society of Chemistry
Creative Commons BY-NC license

Chem. Sci., 2022, Accepted Manuscript

Structural and dynamical determinants of a β-sheet-enriched intermediate involved in amyloid fibrillar assembly of human prion protein

L. Russo, G. Salzano, A. Corvino, E. Bistaffa, F. Moda, L. Celauro, G. D'Abrosca, C. Isernia, D. Milardi, G. Giachin, G. Malgieri, G. Legname and R. Fattorusso, Chem. Sci., 2022, Accepted Manuscript , DOI: 10.1039/D2SC00345G

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