NR ARVA
AU Todorova-Balvay,D.; Simon,S.; Creminon,C.; Grassi,J.; Srikrishnan,T.; Vijayalakshmi,M.A.
TI Copper binding to prion octarepeat peptides, a combined metal chelate affinity and immunochemical approaches
QU Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences 2005 Apr 15; 818(1): 75-82
PT journal article
AB Based on the hypothetical proposal of Sulkowski [E. Sulkowski, FEBS Lett. 307 (2) (1992) 129] for the implication of transition metal ions in the structural changes/oligomerisation of normal cellular prion protein (PrPc) resulting in the pathological isoform (PrPsc), we focused our study on the octarepat domain of this protein which has been supposed to be the metal binding site. We have studied the copper binding to synthetic prion octarepeat peptides (PHGGGWGQ)n (n=1, 3, 6) using metal chelate and size-exclusion modes of chromatographies. This copper binding induces oligomerisation resulting in multiple aggregates. Moreover, heterogeneity of metal bound octarepeat oligomers by ESI-MS has been demonstrated. In addition, anti prion antibodies specific to the octarepeat region were used to discriminate between metal free and copper, nickel and zinc bound hexamer octarepeat peptide. Differential recognition of Cu(II) and Zn(II) bound complexes has been observed which signify differences in exposed epitopes of aggregated peptides.
MH Antigen-Antibody Reactions; Binding Sites; Chelating Agents; Chromatography, Affinity/*methods; Chromatography, Gel; Chromatography, High Pressure Liquid; Copper/*metabolism; Enzyme-Linked Immunosorbent Assay; Nickel/metabolism; Peptides/chemistry/immunology/*metabolism; Prions/chemistry/immunology/*metabolism; Protein Conformation/drug effects; Protein Structure, Quaternary; Spectrometry, Mass, Electrospray Ionization; Zinc/metabolism
AD Laboratoire d'Interactions Moleculaires et de Technologie de Separation (LIMTechS), Centre de Recherche de Royallieu, BP 20.529 Universite de Technologie de Compiegne, F-60205 Compiegne Cedex, France.
SP englisch
PO USA