000144823 001__ 144823
000144823 005__ 20250718160531.0
000144823 0247_ $$2URN$$aurn:nbn:de:hbz:5n-48485
000144823 037__ $$aDZNE-2020-00265
000144823 041__ $$aGerman
000144823 1001_ $$0P:(DE-2719)2810494$$aWüsten, Annick$$b0$$eFirst author$$udzne
000144823 245__ $$aEin zellbasierter Biolumineszenzassay zur Untersuchung der Dimerisierung und Aggregation des Prionproteins$$f - 2017-06-28
000144823 260__ $$aBonn$$c2017
000144823 300__ $$a92 pages : illustrations
000144823 3367_ $$2DataCite$$aOutput Types/Dissertation
000144823 3367_ $$2ORCID$$aDISSERTATION
000144823 3367_ $$2BibTeX$$aPHDTHESIS
000144823 3367_ $$02$$2EndNote$$aThesis
000144823 3367_ $$0PUB:(DE-HGF)11$$2PUB:(DE-HGF)$$aDissertation / PhD Thesis$$bphd$$mphd$$s1752847446_2393
000144823 3367_ $$2DRIVER$$adoctoralThesis
000144823 502__ $$aDissertation, Rheinische Friedrich-Wilhelms-Universität Bonn, 2017$$bDissertation$$cRheinische Friedrich-Wilhelms-Universität Bonn$$d2017
000144823 520__ $$aPrion 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.
000144823 536__ $$0G:(DE-HGF)POF3-342$$a342 - Disease Mechanisms and Model Systems (POF3-342)$$cPOF3-342$$fPOF III$$x0
000144823 8564_ $$uhttps://pub.dzne.de/record/144823/files/DZNE-2020-00265_Restricted.pdf
000144823 8564_ $$uhttps://pub.dzne.de/record/144823/files/DZNE-2020-00265_Restricted.pdf?subformat=pdfa$$xpdfa
000144823 909CO $$ooai:pub.dzne.de:144823$$pVDB
000144823 9101_ $$0I:(DE-588)1065079516$$6P:(DE-2719)2810494$$aDeutsches Zentrum für Neurodegenerative Erkrankungen$$b0$$kDZNE
000144823 9131_ $$0G:(DE-HGF)POF3-342$$1G:(DE-HGF)POF3-340$$2G:(DE-HGF)POF3-300$$3G:(DE-HGF)POF3$$4G:(DE-HGF)POF$$aDE-HGF$$bGesundheit$$lErkrankungen des Nervensystems$$vDisease Mechanisms and Model Systems$$x0
000144823 9141_ $$y2017
000144823 920__ $$lyes
000144823 9201_ $$0I:(DE-2719)1013022$$kAG Tamgüney$$lPrion and prion-like diseases$$x0
000144823 980__ $$aphd
000144823 980__ $$aVDB
000144823 980__ $$aI:(DE-2719)1013022
000144823 980__ $$aUNRESTRICTED