Prion Proteins with Insertion Mutations Have Altered N-terminal Conformation and Increased Ligand Binding Activity and Are More Susceptible to Oxidative Attack

We compared the biochemical properties of a wild type recombinant normal human cellular prion protein, rPrPc, with a recombinant mutant human prion protein that has three additional octapeptide repeats, rPrP8OR. Monoclonal antibodies that are specific for the N terminus of rPrPc react much better wi...

Full description

Saved in:
Bibliographic Details
Published in:The Journal of biological chemistry 2006-04, Vol.281 (16), p.10698-10705
Main Authors: Yin, Shaoman, Yu, Shuiliang, Li, Chaoyang, Wong, Poki, Chang, Binggong, Xiao, Fan, Kang, Shin-Chung, Yan, Huimin, Xiao, Gengfu, Grassi, Jacques, Tien, Po, Sy, Man-Sun
Format: Article
Language:English
Subjects:
Amino Acid Motifs
Antibodies, Monoclonal - chemistry
Calcium - metabolism
Carbon - chemistry
Cations
Copper - chemistry
DNA, Complementary - metabolism
Dose-Response Relationship, Drug
Electrophoresis, Polyacrylamide Gel
Enzyme-Linked Immunosorbent Assay
Epitopes
Glycosaminoglycans - chemistry
Humans
Ligands
Magnetic Resonance Spectroscopy
Models, Genetic
Mutagenesis, Site-Directed
Mutation
Oxygen - chemistry
Oxygen - metabolism
Peptides - chemistry
Prions - chemistry
Prions - metabolism
Protein Binding
Protein Structure, Tertiary
PrPC Proteins - chemistry
Recombinant Proteins - chemistry
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:We compared the biochemical properties of a wild type recombinant normal human cellular prion protein, rPrPc, with a recombinant mutant human prion protein that has three additional octapeptide repeats, rPrP8OR. Monoclonal antibodies that are specific for the N terminus of rPrPc react much better with rPrP8OR than rPrPc, suggesting that the N terminus of rPrP8OR is more exposed and hence more available for antibody binding. The N terminus of PrPc contains a glycosaminoglycan binding motif. Accordingly, rPrP8OR also binds more glycosaminoglycan than rPrPc. In addition, the divalent cation copper modulates the conformations of rPrPc and rPrP8OR differently. When compared with rPrPc, rPrP8OR is also more susceptible to oxidative damage. Furthermore, the abnormalities associated with rPrP8OR are recapitulated, but even more profoundly, in another insertion mutant, which has five extra octapeptide repeats, rPrP10OR. Therefore, insertion mutants appear to share common features, and the degree of abnormality is proportional to the number of insertions. Any of these anomalies may contribute to the pathogenesis of inherited human prion disease.
ISSN:0021-9258
1083-351X
DOI:10.1074/jbc.M511819200