A Bioluminescent Cell Assay to Quantify Prion Protein Dimerization

Sci Rep. 2018 Sep 21;8(1):14178. doi: 10.1038/s41598-018-32581-1.


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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 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
  • Prion Proteins