Home > Publications Database > Ein zellbasierter Biolumineszenzassay zur Untersuchung der Dimerisierung und Aggregation des Prionproteins > print

001     144823
005     20250718160531.0
024 7 _ |a urn:nbn:de:hbz:5n-48485
|2 URN
037 _ _ |a DZNE-2020-00265
041 _ _ |a German
100 1 _ |a Wüsten, Annick
|0 P:(DE-2719)2810494
|b 0
|e First author
|u dzne
245 _ _ |a Ein zellbasierter Biolumineszenzassay zur Untersuchung der Dimerisierung und Aggregation des Prionproteins
|f - 2017-06-28
260 _ _ |a Bonn
|c 2017
300 _ _ |a 92 pages : illustrations
336 7 _ |a Output Types/Dissertation
|2 DataCite
336 7 _ |a DISSERTATION
|2 ORCID
336 7 _ |a PHDTHESIS
|2 BibTeX
336 7 _ |a Thesis
|0 2
|2 EndNote
336 7 _ |a Dissertation / PhD Thesis
|b phd
|m phd
|0 PUB:(DE-HGF)11
|s 1752847446_2393
|2 PUB:(DE-HGF)
336 7 _ |a doctoralThesis
|2 DRIVER
502 _ _ |a Dissertation, Rheinische Friedrich-Wilhelms-Universität Bonn, 2017
|c Rheinische Friedrich-Wilhelms-Universität Bonn
|b Dissertation
|d 2017
520 _ _ |a Prion diseases like Creutzfeldt-Jakob disease (CJD) are fatal neurodegenerative disorders that are triggered by misfolding of the cellular prion protein (PrPC) to an infectious isoform rich in β-sheet structure termed PrPSc. There are evidence that PrPC possibly forms dimers indicating that dimerization of PrPC may be an essential intermediate step for the formation of PrPSc aggregates upon infection.To study PrP dimerization and PrP aggregation upon infection, a bimolecular complementation assay was developed using RK13 cells that lack endogenous PrP expression and stably expressed two fusion constructs between PrP and each half of split Gaussia luciferase. In these cells bioluminescence only occured when PrP dimerized or aggregated and the two split luciferase halves complemented each other.Using a panel of eight antibodies directed against different PrP domains, the central domain of PrPC was identified to be critical for dimerization.Unlike normal brain homogenate infection of RK13 cells with brain homogenates from diseased animals containing six different mouse-adapted prion strains, RML, 22L, ME7, 87V, 79A, and 22A, lead to an increase in bioluminescence. In contrast, treatment of chronically infected but not uninfected RK13 cells with the anti-prion compound quinacrine reduced bioluminescence.Finally, a compound-screening with a compound library including 1650 bioactive compounds based on the bimolecular complementation assay identified JTC-801 as one compound that inhibited PrPC-dimerization. Treatment of ScN2a cells with JTC-801 reduced PrPSc levels in these cells at nanomolar concentrations.Overall, these results showed that PrPC clearly dimerizes in cells and that prion infection led to increased bioluminescence in the newly developed bimolecular complementation assay. The identification of JTC-801 as a bioactive compounds that inhibited PrPC-dimerization and reduced PrPSc-levels in ScN2a cells at nanomolar concentrations suggested that PrPC dimerization may be a critical step in the conversion process of PrPC to PrPSc.
536 _ _ |a 342 - Disease Mechanisms and Model Systems (POF3-342)
|0 G:(DE-HGF)POF3-342
|c POF3-342
|f POF III
|x 0
856 4 _ |u https://pub.dzne.de/record/144823/files/DZNE-2020-00265_Restricted.pdf
856 4 _ |u https://pub.dzne.de/record/144823/files/DZNE-2020-00265_Restricted.pdf?subformat=pdfa
|x pdfa
909 C O |p VDB
|o oai:pub.dzne.de:144823
910 1 _ |a Deutsches Zentrum für Neurodegenerative Erkrankungen
|0 I:(DE-588)1065079516
|k DZNE
|b 0
|6 P:(DE-2719)2810494
913 1 _ |a DE-HGF
|b Gesundheit
|l Erkrankungen des Nervensystems
|1 G:(DE-HGF)POF3-340
|0 G:(DE-HGF)POF3-342
|3 G:(DE-HGF)POF3
|2 G:(DE-HGF)POF3-300
|4 G:(DE-HGF)POF
|v Disease Mechanisms and Model Systems
|x 0
914 1 _ |y 2017
920 _ _ |l yes
920 1 _ |0 I:(DE-2719)1013022
|k AG Tamgüney
|l Prion and prion-like diseases
|x 0
980 _ _ |a phd
980 _ _ |a VDB
980 _ _ |a I:(DE-2719)1013022
980 _ _ |a UNRESTRICTED

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