NR ATIR
AU Petrakis,S.; Sklaviadis,T.K.
TI Isolation and identification of brain proteins that bind to refolded recombinant prion protein
QU International Conference - Prion 2005: Between fundamentals and society's needs - 19.10.-21.10.2005, Congress Center Düsseldorf - Poster Session: Cell Biology of PrPc and PrPsc CELL-49
PT Konferenz-Poster
AB
PrPc, the cellular prion protein, is widely expressed in most tissues, including brain, muscle and the gastrointestinal tract, but its physiological role still remains unclear. During propagation of Transmissible Spongiform Encephalopathies (TSEs), prion protein is converted to the pathological isoform, PrPsc, in a procedure believed to be mediated by as yet unknown host factors. The identification of proteins associated with PrP may provide information about both the biology of prions and the pathogenesis of TSEs.
In the present work, we have identified proteins from normal brain tissue based on their ability to bind to prion protein. Recombinant His-tagged PrP was expressed in E.coli, immobilized and purified on a Ni-NTA column and refolded by gradient removal of the chaotropic agent. Normal brain homogenates were passed over columns with the immobilized refolded PrP. After washes with increasing concentrations of detergents to remove non-PrP binding proteins, prion "partners" were eluted from the column and identified using mass spectroscopy techniques. Following this methodology, we identified PrP-associated proteins participating in the endocytosis pathway, the formation of the membrane and cytosolic skeleton and signal transduction. In future experiments we will examine whether these proteins participate in the PrP conversion mechanism and the development of TSEs.
AD S.Petrakis, T.Sklaviadis, Prion Disease Research Group, Laboratrory of Pharmacology, School of Pharmacy, Aristotle University of Thessaloniki, 54124, Thessaloniki, Greeece
SP englisch
PO Deutschland