NR ATRB
AU Leliveld,S.R.; Dame,R.; Gijs,W.; Korth,C.
TI Expansions in the octarepeat domain of the prion protein effect highly selective binding to prions
QU International Conference - Prion 2005: Between fundamentals and society's needs - 19.10.-21.10.2005, Congress Center Düsseldorf - Poster Session: Structure of PrP and molecular determinants of infectivity STRCT-17
PT Konferenz-Poster
AB
Mutational insertion of additional repeats into the copper-binding N-terminal octarepeat (OR) domain of PrP, residues 60 to 91, has been genetically linked to familial Creutzfeldt-Jakob disease (fCJD) and thereby to de novo generation of prion infectivity. The wild-type, four OR domain is redundant for prion conversion, as its deletion does not significantly hamper propagation of infectivity. To investigate the role of the OR domain in prion de novo generation, we expressed glutathione S-transferase fusion proteins carrying OR domains of varying length (GST-OR) in E. coli. We found that OR domains displayed intriguing differences in their biochemical and biophysical properties depending on the number of ORs present. First, expanded OR domains as occuring in fCJD had the unique ability to bind PrPsc in a tight, detergent-resistant complex. Second, we observed that expanded but not wild-type OR domains were able to form distinct multimeric complexes in a pH-dependent manner. Apart from introducing a novel tool for characterizing PrPsc without the need for ultracentrifugation or protease digestion, we put forward the idea that PrP molecules carrying an expanded OR domain help to recruit and stabilize PrPsc-like conformations in vivo, thus greatly facilitating de novo generation of human prions in fCJD with multiple OR insertions.
Supported by a grant from the Bundesministerium für Bildung und Forschung (BMBF), Germany
AD Rutger Leliveld, Carsten Korth1, University of Düsseldorf, Germany; Remus Dame, Wuite Gijs, Free University Amsterdam, The Netherlands
SP englisch
PO Deutschland