NR ATHE
AU Provansal,M.; Roche,S.; Pastore,M.; Casanova,D.; Windl,O.; Lehmann,S.L.
TI Proteomic analysis of N2a neuroblastoma cells inducible for PrP expression
QU International Conference - Prion 2005: Between fundamentals and society's needs - 19.10.-21.10.2005, Congress Center Düsseldorf - Poster Session: Cell Biology of PrPc and PrPsc CELL-10
PT Konferenz-Poster
AB
The cellular isoform of the prion protein (PrPc) is involved in the appearance and the transmission of transmissible spongiform encephalopathies (TSEs). The exact function of PrPc is still elusive but seems linked in various models with resistance to oxidative stress, neuronal outgrowth and cell survival through specific signal transduction pathways.
Interestingly, only a few molecules have been directly involved in the normal function of the protein, in its cell biology or its conversion into the pathological isoform, PrPsc.
To further investigate the normal function of the protein and progress toward the identification of important factors in TSEs, we decided to use a differential bidimensional (2D) proteomic approach and looked at N2a cells expressing various amounts of PrP after stable transfection with an inducible expression system for PrP (tet-ON) (Windl et al. J Gen Virol. 80:15-21). We prepared 2D gels from at least three protein extracts collected at different time points from cells induced or not induced for PrP expression. After silver staining of the gels we used image analysis (Melanie software from Amersham-Pharmacia) to select more than 30 protein spots which were significantly differentially expressed between the two conditions.
We expect to gain important information regarding PrP function following the identification of the proteins of interest from the gels using peptide mass fingerprinting with MALDI-TOF and MS/MS.
AD Monique Provansal, Stéphane Roche, Manuela Pastore, Danielle Casanova, Sylvain Lehmann, CNRS IGH UPR1142, Montpellier, France; Otto Windl, VLA Weybridge, UK
SP englisch
PO Deutschland