NR AOSB
AU Eiden,M.; Garner,K.; Groschup,M.H.
TI Conversion of recombinant prion protein in a cell-free system
QU International Conference - Prion diseases: from basic research to intervention concepts - TSE-Forum, 08.10.-10.10.2003, Gasteig, München - Poster session - BR-30
PT Konferenz-Poster
AB
Transmissible spongiform encephalopathies are fatal neurodegenerative diseases in humans and animals and are characterized by the accumulation of a disease-specific, proteinase K (PK) resistant isoform of prion protein, designated PrPsc, in the central nervous system. PrPsc is generated posttranslationally by a conversion reaction from the normal, protease-sensitive isoform of PrPc.
This conversion process can be imitated in a cell-free conversion reaction by adding PrPsc (purified from several animal sources) to recombinant PrPc. PrPc converts subsequently into a PK-resistant form PrPres. In earlier publications, cell-free assays were carried out with PrPc purified from mammalian cell culture or baculovirus-infected cells. Here we present a cell-free system utilizing PrPc produced in E. coli. Moreover, we used a non-radioactive visualization system for detection of newly formed PrPres which is based on a PrP species specific monoclonal antibody in combination with enhanced chemiluminescense.
This system constitutes a simple and well-defined method for cell-free conversion of prion proteins with potential to replace radiolabeled PrPc which has to be elaboratorily purified from tissue culture cell homogenates. By use of this new assay format, high throuput screens for inhibitors of PrPres formation become possible now which may constitute effective drugs against prion diseases.
AD M. Eiden, K. Garner, M.H. Groschup, Federal Research Centre for Virus Diseases of Animals, Germany
SP englisch
PO Deutschland