Author/Editor     Dvorakova, Eva; Vranac, Tanja; Janouskova, Olga; Černilec, Maja; Koren, Simon; Lukan, Anja; Nováková, Jana; Matej, Radoslav; Holada, Karel; Čurin-Šerbec, Vladka
Title     Detection of the GPI-anchorless prion protein fragment PrP226* in human brain
Type     članek
Vol. and No.     Letnik 13
Publication year     2013
ISSN     1471-2377 - BMC neurology
Language     eng
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 spongi form 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).ConclusionsIn 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.
Keywords     transmissible spongiform encephalopathies
Creutzfeldt-Jakob disease
immunology
prenosljive spongiformne encefalopatije
Creutzfeldt-Jakobova bolezen
imunologija