NR AWQS
AU Xanthopoulos,K.; Polymenidou,M.; Sklaviadis,T.K.
TI Comparative inter-species carbohydrate characterization of PrPsc by lectins
QU International Conference - Prion 2006: Strategies, advances and trends towards protection of society - 3.10.-6.10.2006, Torino, Italy, Lingotto Conference Centre - Poster sessions PR-36
PT Konferenz-Poster
AB The key event in transmissible spongiform encephalopathies (TSEs) is the conversion of the soluble, protease-sensitive glycosylated prion protein (PrPc) to an abnormally structured, aggregated and protease-resistant isoform (PrPsc). Both PrP isoforms bear two glycosylation sites, none of which is necessarily occupied, and thus in a typical western blot with an anti-PrP antibody three distinct bands appear, each corresponding to the di- the mono- or the unglycosylated form of the protein. The intensity and the electrophoretic mobility of each of the three bands is characteristic of each TSE type and has been used to discriminate between the various TSE strains and types. In the present study we examined the possibility that in addition to quantitative differences between the amounts of each glycoform, there are also variations in the sugars carried by PrPsc from different species, using lectins. Lectins are multimeric proteins, which can bind with high affinity to specific sugar moieties. PrPsc was purified from various TSE affected samples using a protocol based on differential guanidinium precipitation over a sucrose cushion, followed by proteinase-K treatment and salt precipitation. PrPsc was then analyzed by SDS-PAGE, electrotransferred on PVDF membranes and western blotted with a panel of biotinylated lectins and monoclonal anti-PrP antibodies. From this study were identified two plant lectins, namely Datura stramonium Lectin (DSL) and Ricinus communis Agglutinin I (RCAI), that bind PrPsc with high affinity. Furthermore, it was found that only a subpopulation of the PrPsc population is recognized by these lectins and that there are differences in the affinity with which these lectins recognize the PrPsc glycoforms in the various TSE types. Lectin staining of PrPsc could prove to be a useful tool for studying the effect of the host and the TSE strain based on glycotype profile and discriminating between the various types of TSEs.
AD K. Xanthopoulos, T. Sklaviadis: Laboratory of Pharmacology, Department of Pharmacy, School of Health Sciences, Aristotle University of Thessaloniki, Thessaloniki 54124, Greece and Centre for Research and Technology-Hellas, Institute of Agrobiotechnology, CERTH/INA 6th km Charilaou Thermi, 57001 Thessaloniki, Greece; M. Polymenidou: Institute of Neuropathology, University Hospital of Zürich, Schmelzbergstrasse 12, CH-8091 Zürich, Switzerland. E-mail: K. Xanthopoulos: xantho@pharm.auth.gr
SP englisch
PO Italien
EA Poster, Übersicht, Ausschnitt 1, Ausschnitt 2, Ausschnitt 3