Photo-induced crosslinking of prion protein oligomers and prions

Prion diseases are caused by a unique type of infectious agent, which is thought to consist of a misfolded β-sheeted form of the α-helical cellular prion protein (PrPC). This misfolded isoform (PrPSc) tends to form insoluble amyloid-like aggregates, impeding classical structural analysis by X-ray cr...

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Bibliographic Details
Published in:Amyloid 2006-06, Vol.13 (2), p.67-77
Main Authors: Piening, Niklas, Weber, Petra, Högen, Tobias, Beekes, Michael, Kretzschmar, Hans, Giese, Armin
Format: Article
Language:English
Subjects:
amyloid
Amyloid - chemistry
Amyloid - genetics
Amyloid - metabolism
Animals
Brain Chemistry
crosslinking
dementia
Humans
Mice
misfolding
neurodegeneration
oligomer
Photochemistry - methods
PICUP
Prion Diseases - genetics
Prion Diseases - metabolism
prion protein
Protein Folding
PrPC Proteins - chemistry
PrPC Proteins - genetics
PrPC Proteins - metabolism
PrPSc Proteins - chemistry
PrPSc Proteins - genetics
PrPSc Proteins - metabolism
Recombinant Proteins - chemistry
Recombinant Proteins - genetics
seeding
structure
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Summary:Prion diseases are caused by a unique type of infectious agent, which is thought to consist of a misfolded β-sheeted form of the α-helical cellular prion protein (PrPC). This misfolded isoform (PrPSc) tends to form insoluble amyloid-like aggregates, impeding classical structural analysis by X-ray crystallography or NMR. Intermolecular crosslinking may provide a means of stabilizing notoriously elusive oligomers for further analysis and may be used for analyzing aggregate architecture by characterising intermolecular contact sites. Using a photo-induced crosslinking method (PICUP), aggregates of recombinant PrP (rPrP) and PrPSc were linked at interacting surfaces via amino acid side chains. The degree of crosslinking within PrP aggregates was adjustable using varying light intensities and could efficiently be monitored by fluorescence correlation spectroscopy. Specific intermolecular crosslinking of PrPSc molecules was achieved even in crude brain homogenate. Functional studies showed that stabilized aggregates of rPrP did not loose their capacity to induce further protein aggregation and crosslinking of PrPSc did not alter significantly the level of infectivity, indicating that photo-induced covalent linkage of PrPSc does not destruct surfaces important for prion propagation.
ISSN:1350-6129
1744-2818
DOI:10.1080/13506120600722498