NR ATIJ
AU Ramljak,S.; Asif,A.R.; Armstrong,V.W.; Weise,J.; Buschmann,A.; Bodemer,W.; Zerr,I.
TI Protein profiling in murine brain PrP0/0 cells lacking the cellular prion protein gene (PRNP) vs. PrP0/0 cells transiently transfected with human PRNP
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-41
PT Konferenz-Poster
AB
The cellular isoform of prion protein (PrPc) is widely distributed in neural and non-neural tissues with the highest level at the cell surface of neurons in the central nervous system (CNS). The abundance of PrPc in the CNS argues for an important physiological function. The goal of the present study was to gain insight into its physiological function(s) by comparing protein profiles of cells in the absence or presence of PrPc. Therefore, we analysed the protein profiles of immortalised prion protein gene deficient (PrP0/0) mouse brain cells vs. PrP0/0 cells transiently transfected with human PrPc. Protein analyses were performed with two dimensional gel electrophoresis (2-DE) and electrospray ionization time of flight mass spectrometry (ESI-TOF-MS).
By comparing protein patterns of PrP0/0 cells and PrPc - transfected null cells we detected two differentially expressed protein spots which were identified as Annexin II (38 kDa, pI=7.55) and Annexin V (36 kDa, pI=4.83) by subsequent peptide sequencing analysis. Western blot analysis confirmed Annexin V to be more abundant in PrPc transfected cells vs. PrP0/0 cells. These results suggest a possible PrP-dependent regulation/biosynthesis of Annexin V. We also detected an enhanced abundance of the Annexin V protein in HEK 293 cells transiently overexpressing PRNP compared to 293 cells transfected with empty vector.
Preliminary data on PrPc and Annexin V may point to common interaction-mechanisms in transfected PrP null cells. Ongoing experiments are elucidating the functional relevance of these in vitro results by investigating Annexin V - PrPc interaction in PrP0/0 and PrPc expressing mice under physiological conditions and after focal cerebral ischemia.
AD Sanja Ramljak, Jens Weise, Inga Zerr, Dept. of Neurology, Georg August University, Göttingen, Germany; Abdul Rahman Asif, Victor W. Armstrong, Dept. of Clinical Chemistry, Georg August University, Göttingen, Germany; Anne Buschmann, Friedrich Loeffler Institut, INEID, Greifswald-Insel Riems, Germany; Walter Bodemer, Dept. of Infectious Pathology, German Primate Center, Göttingen, Germany
SP englisch
PO Deutschland