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BMC Neurology
Published in: BMC Neurology 1/2013

Open Access 01-12-2013 | Research article

Detection of the GPI-anchorless prion protein fragment PrP226* in human brain

Authors: Eva Dvorakova, Tanja Vranac, Olga Janouskova, Maja Černilec, Simon Koren, Anja Lukan, Jana Nováková, Radoslav Matej, Karel Holada, Vladka Čurin Šerbec

Published in: BMC Neurology | Issue 1/2013

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Abstract

Background

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.

Methods

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.

Results

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).

Conclusions

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.
Appendix
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Metadata
Title
Detection of the GPI-anchorless prion protein fragment PrP226* in human brain
Authors
Eva Dvorakova
Tanja Vranac
Olga Janouskova
Maja Černilec
Simon Koren
Anja Lukan
Jana Nováková
Radoslav Matej
Karel Holada
Vladka Čurin Šerbec
Publication date
01-12-2013
Publisher
BioMed Central
Published in
BMC Neurology / Issue 1/2013
Electronic ISSN: 1471-2377
DOI
https://doi.org/10.1186/1471-2377-13-126

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