NR AXYM
AU Watts,J.C.; Ng,V.; Horne,P.; Schmitt-Ulms,G.; Fraser,P.E.; Westaway,D.
TI Biochemical Properties of Wild Type Shadoo Protein from Mice
QU International Conference - Prion 2007 (26.-28.9.2007) Edinburgh International Conference Centre, Edinburgh, Scotland, UK - Book of Abstracts: Pathology and Pathogenesis P03.119
IA http://www.prion2007.com/pdf/Prion Book of Abstracts.pdf
PT Konferenz-Poster
AB Shadoo is a notional PrP-like protein encoded by the SPRN gene. Using a variety of antibodies we demonstrate that Shadoo is expressed as a glycosylated, GPI-anchored protein in the mature CNS. Shadoo undergoes endoproteolytic processing both in cultured cells and in the adult mouse brain to generate a C-terminal fragment somewhat analogous to the PrP "C1" cleavage product. The domain architecture of wild-type Shadoo, which comprises a signal peptide, a charged region, a region with homology to PrP's hydrophobic region and a short C-terminal region, is notable in resembling the domain architecture of PrP23-144: the latter molecule is synthesized in patients bearing a PRNP Y145Stop mutation and is prone to form amyloid fibrils in vitro and in vivo. Consequently, we have evaluated the ability of wild-type mouse Shadoo to form fibrils in vitro. Full-length recombinant mouse Shadoo was produced in E. coli from a variety of recombinant plasmids and was purified to homogeneity. While soluble in aqueous solution and natively unstructured, upon incubation at 37° recombinant Shadoo formed both fibrillar and pre-fibrillar structures. Furthermore, since (i) homology to PrP lies within the hydrophobic domain that is misfolded in PrPsc, (ii) certain areas of Shadoo expression in the mouse brain correspond to early target sites during prion disease, and (iii) GPI-cleaved Shadoo and PrPsc might interact in the parenchyma of the CNS, we have considered whether wild-type Shadoo is modulated by - or may modulate - PrP misfolding. Based upon the ability of wild-type Shadoo to adopt ß-structure we believe that Shadoo is uniquely plausible as a modulator of prion disease pathogenesis.
AD J.C. Watts, V. Ng, P. Horne, G. Schmitt-Ulms, P.E. Fraser, Centre for Research in Neurodegenerative Disease, University of Toronto, Canada; D. Westaway, Alberta Centre for Prions and Protein Folding Diseases, University of Alberta, Canada
SP englisch
PO Schottland