Detection of the GPI-anchorless prion protein fragment PrP226* in human brain
- Equal contributors
1 Institute of Immunology and Microbiology, 1st Faculty of Medicine, Charles University in Prague, Studnickova 7, 128 20 Prague 2, Czech Republic
2 Department for Production of Diagnostic Reagents and Research, Blood Transfusion Centre of Slovenia, Šlajmerjeva 6, 1000 Ljubljana, Slovenia
3 Department of Pathology and Molecular Medicine, Thomayer Hospital, Vídeňská 800, 140 59 Prague 4, Czech Republic
4 Current address: Institute of Macromolecular Chemistry v.v.i., AS CR, Heyrovsky square 2, 162 00 Prague 6, Czech Republic
5 Current address: Omega, d.o.o., Dolinškova 8, 1000 Ljubljana, Slovenia
BMC Neurology 2013, 13:126 doi:10.1186/1471-2377-13-126Published: 25 September 2013
The accumulation of the misfolded forms of cellular prion protein, i.e. prions (PrPSc), in the brain is one of the crucial characteristics of fatal neurodegenerative disorders, called transmissible spongiform encephalopathies (TSEs). Cellular prion protein is normally linked to the cell surface by the glycosylphosphatidylinositol (GPI) anchor. There is accumulating evidence that the GPI-anchorless prion protein may act as an accelerator of formation and propagation of prions. In the TSE affected human brain we have previously discovered a novel GPI-anchorless prion protein fragment, named PrP226*, which ends with the tyrosine 226. This fragment can be labeled specifically by the monoclonal antibody V5B2.
We developed a DELFIA based assay for quick and sensitive detection of the PrP226* fragment in human brain tissue homogenates. By calculating the ratio between the signals of native (N) and denatured (D) samples applied to the assay we were able to observe significant difference between 24 TSE affected brains and 10 control brains. The presence of PrP226* in brain tissue was confirmed by western blot.
Our results demonstrate that PrP226* is present in small quantities in healthy human brain, whereas in degenerated brain it accumulates in prion aggregates, proportionally to PrPSc. Samples with high D/N ratio generally comprised more proteinase K resistant PrP, while no correlation was found between the quantity of PrP226* and standard classification of Creutzfeldt-Jakob disease (CJD).
In the present study we show that the PrP226* fragment accumulates in prion aggregates and after being released from them by a denaturation procedure, could serve as a proteinase K digestion independent biomarker for human TSEs. The PrP226* assay described in this paper offers a tool to follow and study this unique anchorless PrP fragment in various parts of human brain and possibly also in other tissues and body fluids.