Impact of N-glycosylation site variants during human PrP aggregation and fibril nucleation.

Authors:
Rajesh Mishra
Rajesh Mishra
Linköping University
Sweden
Mathias Elgland
Mathias Elgland
Linköping University
Cape Town | South Africa
Afshan Begum
Afshan Begum
University of Karachi
Pakistan
Timmy Fyrner
Timmy Fyrner
Linköping University
Sweden
Peter Konradsson
Peter Konradsson
Linköping University
Sweden
Per Hammarstrom
Per Hammarstrom
Linköping University

Biochim Biophys Acta Proteins Proteom 2019 Mar 30. Epub 2019 Mar 30.

IFM-Department of Physics, Chemistry and Biology, Linköping University, Linköping, Sweden. Electronic address:

Misfolding and aggregation of the human prion protein (PrP) cause neurodegenerative transmissible spongiform encephalopathies such as Creutzfeldt-Jakob disease. Mature native PrP is composed of 209 residues and is folded into a C-terminal globular domain (residues 125-209) comprising a small two-stranded β-sheet and three α-helices. The N-terminal domain (residues 23-124) is intrinsically disordered. Expression of truncated PrP (residues 90-231) is sufficient to cause prion disease and residues 90/100-231 is comprising the amyloid-like fibril core of misfolded infectious PrP. During PrP fibril formation under native conditions in vitro, the disordered N-terminal domain slows down fibril formation likely due to a mechanism of initial aggregation forming morphologically disordered aggregates. The morphological disordered aggregate is a transient phase. Nucleation of fibrils occurs from this initial aggregate. The aggregate phase is largely circumvented by seeding with preformed PrP fibrils. In vivo PrP is N-glycosylated at positions Asn181 and Asn197. Little is known about the importance of these positions and their glycans for PrP stability, aggregation and fibril formation. We have in this study taken a step towards that goal by mutating residues 181 and 197 for cysteines to study the positional impact on these processes. We have further by organic synthetic chemistry and chemical modification generated synthetic glycosylations in these positions. Our data shows that residue 181 when mutated to a cysteine is a key residue for self-chaperoning, rendering a trap in the initial aggregate preventing conformational changes towards amyloid fibril formation. Position 197 is less involved in the aggregate trapping and is more geared towards β-sheet structure conversion within amyloid fibrils. As expected, synthetic glycosylated 197 is less affected towards fibril formation compared to glycosylated 181. Our data are rather compatible with the parallel in-register intermolecular β-sheet model structure of the PrP90-231 fibril and sheds light on the misfolding transitions of PrP in vitro. We hypothesize that glycosylation of position 181 is a key site for prion strain differentiation in vivo.

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http://dx.doi.org/10.1016/j.bbapap.2019.03.010DOI Listing

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March 2019
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