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@ARTICLE{Wsten:140214,
author = {Wüsten, Annick and Pasham Parameshwar, Reddy and Smiyakin,
Andrej and Bernis, Maria Eugenia and Tamgüney, Gültekin},
title = {{A} {B}ioluminescent {C}ell {A}ssay to {Q}uantify {P}rion
{P}rotein {D}imerization.},
journal = {Scientific reports},
volume = {8},
number = {1},
issn = {2045-2322},
address = {[London]},
publisher = {Macmillan Publishers Limited, part of Springer Nature},
reportid = {DZNE-2020-06536},
pages = {14178},
year = {2018},
abstract = {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.},
keywords = {Animals / Biological Assay: methods / Cations, Divalent:
metabolism / Cell Line / Cell Line, Tumor /
Creutzfeldt-Jakob Syndrome: metabolism / Deer / Dimerization
/ Humans / Luminescent Measurements: methods / Mice / Prion
Proteins: metabolism / Protein Folding / Rabbits / Scrapie:
metabolism / Sheep / Wasting Disease, Chronic: metabolism /
Cations, Divalent (NLM Chemicals) / Prion Proteins (NLM
Chemicals)},
cin = {AG Tamgüney 2},
ddc = {600},
cid = {I:(DE-2719)1013022},
pnm = {342 - Disease Mechanisms and Model Systems (POF3-342)},
pid = {G:(DE-HGF)POF3-342},
typ = {PUB:(DE-HGF)16},
pubmed = {pmid:30242186},
pmc = {pmc:PMC6155003},
doi = {10.1038/s41598-018-32581-1},
url = {https://pub.dzne.de/record/140214},
}