NR AXQY
AU Lawson,V.A.; Lumicisi,B.; Machalek,D.; Gouramanis,K.; Klemm,H.; Masters,C.L.; Collins,S.J.; Hill,A.F.
TI How Do Glycosaminoglycans Promote the Propagation of Prions?
QU International Conference - Prion 2007 (26.-28.9.2007) Edinburgh International Conference Centre, Edinburgh, Scotland, UK - Book of Abstracts: Protein Misfolding P01.24
IA http://www.prion2007.com/pdf/Prion Book of Abstracts.pdf
PT Konferenz-Poster
AB
Background: It is recognised that ancillary factors may contribute to the pathogenesis and transmission of prion diseases. Of particular interest have been polyanions, including nucleic acids and glycosaminoglycans (GAGs). However the exact nature of this interaction is poorly understood.
Aim: To elucidate the molecular basis of the GAG prion protein (PrP) interaction.
Methods: We are using a cell free model of protease resistant PrP (PrPres) formation to elucidate the role of GAGs in prion disease. This model, the conversion activity assay (CAA), can be used to generate PrPres from brain homogenate or cell lysate derived PrPc and is driven by a PrPsc seed derived from prion infected tissue. The CAA enables the study of the role of GAGs in the formation of PrPres through degradation/inhibition of endogenous GAGs or disruption of the GAG/PrP interaction. To further characterise the GAG PrP interaction we have modified GAG expression in a rabbit kidney cell line (RK13) expressing exogenous mouse PrP (moRK13). This system is also being used to express mutant forms of PrPc to further investigate the molecular basis of GAG mediated PrPres formation.
Results: Conversion activity in the CAA is disrupted by conditions that disrupt the electrostatic interaction typically associated with a GAG protein interaction. These conditions also disrupt the ability of brain and cell lysate derived PrP to bind a purified source of GAGs. Depletion of endogenous GAGs present in brain homogenates used as a source of PrP in the CAA implicates GAGs in PrPres formation. The molecular basis of this effect is being further investigated using lysates of moRK13 cells in which GAG expression has been modified.
Discussion: GAG analogs and polyanions have shown great potential as anti-prion therapeutics. In this study we are seeking to elucidate the molecular basis of this interaction with the aim of developing effective and targeted therapeutics.
AD V.A. Lawson, B. Lumicisi, D. Machalek, K. Gouramanis, H. Klemm, S.J. Collins, The University of Melbourne, Department of Pathology, Australia; C.L. Masters, Mental Health Research Institute of Victoria, Australia; A.F. Hill, Bio 21 Molecular Science and Biotechnology Institute, Department of Biochemistry and Molecular Biology, Australia
SP englisch
PO Schottland