Rapid folding of the prion protein captured by pressure-jump.
The conversion of the cellular form of the prion protein (PrP(C)) to an altered disease state, generally denoted as scrapie isoform (PrP(Sc)), appears to be a crucial molecular event in prion diseases. The details of this conformational transition are not fully understood, but it is perceived that they are associated with misfolding of PrP or its incapacity to maintain the native fold during its cell cycle. Here we present a tryptophan mutant of PrP (F198W), which has enhanced fluorescence sensitivity to unfolding/refolding transitions. Equilibrium folding was studied by circular dichroism and fluorescence. Pressure-jump experiments were successfully applied to reveal rapid submillisecond folding events of PrP at temperatures not accessed before.
Jenkins
David C
DC
Department of Biological Sciences, University of Warwick, Gibbet Hill Road, Coventry CV4 7AL, UK.
Pearson
David S
DS
Department of Biosciences, University of Kent, Canterbury, Kent CT2 7NJ, UK.
Harvey
Andrew
A
Department of Biological Sciences, University of Warwick, Gibbet Hill Road, Coventry CV4 7AL, UK.
Sylvester
Ian D
ID
Department of Biological Sciences, University of Warwick, Gibbet Hill Road, Coventry CV4 7AL, UK.
Geeves
Michael A
MA
Department of Biosciences, University of Kent, Canterbury, Kent CT2 7NJ, UK.
Pinheiro
Teresa J T
TJT
Department of Biological Sciences, University of Warwick, Gibbet Hill Road, Coventry CV4 7AL, UK.
D000818:Animals; D002942:Circular Dichroism; D006224:Cricetinae; D007700:Kinetics; D008647:Mesocricetus; D009154:Mutation; D010446:Peptide Fragments / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D011312:Pressure; D011328:Prions / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D011489:Protein Denaturation; D017510:Protein Folding; D020673:Protein Renaturation; D055550:Protein Stability; D017434:Protein Structure, Tertiary; D013050:Spectrometry, Fluorescence; D013696:Temperature; D013816:Thermodynamics
D010446:Peptide Fragments; D011328:Prions