TY  - JOUR
AU  - Wüsten, Annick
AU  - Pasham Parameshwar, Reddy
AU  - Smiyakin, Andrej
AU  - Bernis, Maria Eugenia
AU  - Tamgüney, Gültekin
TI  - A Bioluminescent Cell Assay to Quantify Prion Protein Dimerization.
JO  - Scientific reports
VL  - 8
IS  - 1
SN  - 2045-2322
CY  - [London]
PB  - Macmillan Publishers Limited, part of Springer Nature
M1  - DZNE-2020-06536
SP  - 14178
PY  - 2018
AB  - 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.
KW  - Animals
KW  - Biological Assay: methods
KW  - Cations, Divalent: metabolism
KW  - Cell Line
KW  - Cell Line, Tumor
KW  - Creutzfeldt-Jakob Syndrome: metabolism
KW  - Deer
KW  - Dimerization
KW  - Humans
KW  - Luminescent Measurements: methods
KW  - Mice
KW  - Prion Proteins: metabolism
KW  - Protein Folding
KW  - Rabbits
KW  - Scrapie: metabolism
KW  - Sheep
KW  - Wasting Disease, Chronic: metabolism
KW  - Cations, Divalent (NLM Chemicals)
KW  - Prion Proteins (NLM Chemicals)
LB  - PUB:(DE-HGF)16
C6  - pmid:30242186
C2  - pmc:PMC6155003
DO  - DOI:10.1038/s41598-018-32581-1
UR  - https://pub.dzne.de/record/140214
ER  -