%0 Journal Article
%A Wüsten, Annick
%A Pasham Parameshwar, Reddy
%A Smiyakin, Andrej
%A Bernis, Maria Eugenia
%A Tamgüney, Gültekin
%T A Bioluminescent Cell Assay to Quantify Prion Protein Dimerization.
%J Scientific reports
%V 8
%N 1
%@ 2045-2322
%C [London]
%I Macmillan Publishers Limited, part of Springer Nature
%M DZNE-2020-06536
%P 14178
%D 2018
%X The prion protein (PrP) is a cell surface protein that in disease misfolds and becomes infectious causing Creutzfeldt-Jakob disease in humans, scrapie in sheep, and chronic wasting disease in deer and elk. Little is known regarding the dimerization of PrP and its role in disease. We developed a bioluminescent prion assay (BPA) to quantify PrP dimerization by bimolecular complementation of split Gaussia luciferase (GLuc) halves that are each fused to PrP. Fusion constructs between PrP and N- and C-terminal GLuc halves were expressed on the surface of RK13 cells (RK13-DC cells) and dimerized to yield a bioluminescent signal that was decreased in the presence of eight different antibodies to PrP. Dimerization of PrP was independent of divalent cations and was induced under stress. Challenge of RK13-DC cells with seven different prion strains did not lead to detectable infection but was measurable by bioluminescence. Finally, we used BPA to screen a compound library for compounds inhibiting PrP dimerization. One of the most potent compounds to inhibit PrP dimerization was JTC-801, which also inhibited prion replication in RML-infected ScN2a and SMB cells with an EC50 of 370 nM and 220 nM, respectively. We show here that BPA is a versatile tool to study prion biology and to identify anti-prion compounds.
%K Animals
%K Biological Assay: methods
%K Cations, Divalent: metabolism
%K Cell Line
%K Cell Line, Tumor
%K Creutzfeldt-Jakob Syndrome: metabolism
%K Deer
%K Dimerization
%K Humans
%K Luminescent Measurements: methods
%K Mice
%K Prion Proteins: metabolism
%K Protein Folding
%K Rabbits
%K Scrapie: metabolism
%K Sheep
%K Wasting Disease, Chronic: metabolism
%K Cations, Divalent (NLM Chemicals)
%K Prion Proteins (NLM Chemicals)
%F PUB:(DE-HGF)16
%9 Journal Article
%$ pmid:30242186
%2 pmc:PMC6155003
%R 10.1038/s41598-018-32581-1
%U https://pub.dzne.de/record/140214