A Bioluminescent Cell Assay to Quantify Prion Protein Dimerization
| العنوان: | A Bioluminescent Cell Assay to Quantify Prion Protein Dimerization |
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| المؤلفون: | Gültekin Tamgüney, Pasham Parameshwar Reddy, Andrej Smiyakin, Katharina Annick Wüsten, Maria Eugenia Bernis |
| المصدر: | Scientific Reports, Vol 8, Iss 1, Pp 1-14 (2018) Scientific reports 8(1), 14178 (2018). doi:10.1038/s41598-018-32581-1 |
| بيانات النشر: | Nature Publishing Group, 2018. |
| سنة النشر: | 2018 |
| مصطلحات موضوعية: | 0301 basic medicine, Protein Folding, animal diseases, Cell, lcsh:Medicine, Scrapie, Creutzfeldt-Jakob Syndrome, Mice, 0302 clinical medicine, metabolism [Creutzfeldt-Jakob Syndrome], lcsh:Science, Multidisciplinary, biology, Chemistry, Complementation, medicine.anatomical_structure, Biochemistry, Wasting Disease, Chronic, methods [Biological Assay], Biological Assay, Protein folding, Rabbits, Antibody, Dimerization, metabolism [Wasting Disease, Chronic], Cations, Divalent, Prion Proteins, Cell Line, methods [Luminescent Measurements], 03 medical and health sciences, Gaussia, metabolism [Prion Proteins], Cell Line, Tumor, metabolism [Cations, Divalent], metabolism [Scrapie], medicine, Animals, Humans, Luciferase, Sheep, Deer, lcsh:R, biology.organism_classification, nervous system diseases, 030104 developmental biology, Cell culture, Luminescent Measurements, biology.protein, lcsh:Q, ddc:600, 030217 neurology & neurosurgery |
| الوصف: | 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. |
| اللغة: | English |
| تدمد: | 2045-2322 |
| الوصول الحر: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::5bf2ace83d15687580670a1206f4e308Test http://link.springer.com/article/10.1038/s41598-018-32581-1Test |
| حقوق: | OPEN |
| رقم الانضمام: | edsair.doi.dedup.....5bf2ace83d15687580670a1206f4e308 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 20452322 |
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