| Autor: |
O'Sullivan DB; School of Biological and Chemical Sciences, Queen Mary, University of London, Mile End Road, London E1 4NS, UK., Jones CE, Abdelraheim SR, Thompsett AR, Brazier MW, Toms H, Brown DR, Viles JH |
| Jazyk: |
angličtina |
| Zdroj: |
The Biochemical journal [Biochem J] 2007 Jan 15; Vol. 401 (2), pp. 533-40. |
| DOI: |
10.1042/BJ20060668 |
| Abstrakt: |
Prion diseases are associated with the misfolding of the PrP (prion protein) from a largely alpha-helical isoform to a beta-sheet-rich oligomer. CD has shown that lowering the pH to 4 under mildly denaturing conditions causes recombinant PrP to convert from an alpha-helical protein into one that contains a high proportion of beta-sheet-like conformation. In the present study, we characterize this soluble pH 4 folding intermediate using NMR. (15)N-HSQC (heteronuclear single-quantum correlation) studies with mPrP (mouse PrP)-(23-231) show that a total of 150 dispersed amide signals are resolved in the native form, whereas only 65 amide signals with little chemical shift dispersion are observable in the pH 4 form. Three-dimensional (15)N-HSQC-TOCSY and NOESY spectra indicate that the observable residues are all assigned to amino acids in the N-terminus: residues 23-118. (15)N transverse relaxation measurements indicate that these N-terminal residues are highly flexible with additional fast motions. These observations are confirmed via the use of truncated mPrP-(112-231), which shows only 16 (15)N-HSQC amide peaks at pH 4. The loss of signals from the C-terminus can be attributed to line broadening due to an increase in the molecular size of the oligomer or exchange broadening in a molten-globule state. |
| Databáze: |
MEDLINE |
| Externí odkaz: |
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