A role for His155 in binding of human prion peptide144-167 to immobilised prion protein.
The interactions and conformational changes that lead to the conversion of the normal prion protein (PrP(c)) to its pathogenic form, PrP(sc), are still being elucidated. Using Surface Plasma Resonance (SPR), we provide evidence that a synthetic peptide (PrP(144-167)) corresponding to residues comprising the alpha helix 1-beta strand 2 domain of PrP(c) is able to interact and bind to immobilised recombinant human PrP (rHuPrP) in a dose-dependent manner. The interaction is pH dependent with an increase in binding observed as the pH is lowered, particularly between pH 6.5 and pH 5.5 suggesting a specific role for His(155) in the interaction, confirmed by covalent modification of this residue in the peptide with diethylpyrocarbonate (DEPC). Circular dichroism analysis of PrP(144-167) revealed no secondary structure motifs across the pH range investigated. Possible pH related structural changes of immobilised rHuPrP are also discussed with regard to the increased affinity for PrP(144-167).
Hesp
J Richard
JR
Health Protection Agency, Porton Down, Salisbury, Wiltshire SP4 0JG, UK. richard.hesp@hpa.org.uk
Raven
Neil D H
ND
Sutton
J Mark
JM
D020816:Amino Acid Motifs; D002942:Circular Dichroism; D004047:Diethyl Pyrocarbonate / Q000737:chemistry; D004305:Dose-Response Relationship, Drug; D006639:Histidine / Q000737:chemistry*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D011328:Prions / Q000737:chemistry*; D011485:Protein Binding; D011487:Protein Conformation; D017510:Protein Folding; D017433:Protein Structure, Secondary; D011994:Recombinant Proteins / Q000737:chemistry; D020349:Surface Plasmon Resonance
D011328:Prions; D011994:Recombinant Proteins; D006639:Histidine; D004047:Diethyl Pyrocarbonate