NR AXRR
AU Lund,C.; Olsen,C.M.; Tveit,H.; Tranulis,M.A.
TI The Prion Protein 3F4 Epitope Revisited: Characterization of the Minimal Epitope and its use as a Molecular Tag
QU International Conference - Prion 2007 (26.-28.9.2007) Edinburgh International Conference Centre, Edinburgh, Scotland, UK - Book of Abstracts: Protein Misfolding P01.22
IA http://www.prion2007.com/pdf/Prion Book of Abstracts.pdf
PT Konferenz-Poster
AB
The commonly used monoclonal antibody (MAb) 3F4, has for nearly two decades contributed significantly to our understanding of the normal cell biology of the prion protein (PrP), as well as the disease related abnormalities occurring in prion diseases.
The 3F4 antibody binds to human, hamster and feline PrP with high affinity, requiring two Met residues at positions corresponding to 109 and 112 in the human PrP. Species, that lack one of the Met residues, like cattle and sheep, or both, like rat and mouse, do not react with the 3F4 antibody.
Previous observations have led to the commonly accepted notion that the 3F4 epitope consists of a tetra-peptide Met-Lys-His-Met, encompassing residues 109-112 in human PrP. Here, we have characterized the epitope by studying its binding to synthetic peptides and by analysis of mutated ovine PrP::GFP constructs expressed in the murine neuroblastoma cell line N2a. We found the 3F4 epitope to consist of the hepta-peptide Lys-Thr-Asn-Met-Lys-His-Met (106-112), where the Lys 109 is critically important for 3F4 binding. We also demonstrate that the hepta-peptide constituting the minimal 3F4 epitope can be used as a discrete, moveable high-affinity tag, which might be useful in applications beyond prion research.
AD C. Lund, C.M. Olsen, M.A. Tranulis, Norwegian School of Veterinary Sciences, Department of Molecular Biosciences, Norway; H. Tveit, University of Oslo, Norway
SP englisch
PO Schottland