Dynamics of a truncated prion protein, PrP(113–231), from 15N NMR relaxation: Order parameters calculated and slow conformational fluctuations localized to a distinct region

Abstract

Prion diseases are associated with the misfolding of the prion protein (PrP^C) from a largely α‐helical isoform to a β‐sheet rich oligomer (PrP^Sc). Flexibility of the polypeptide could contribute to the ability of PrP^C to undergo the conformational rearrangement during PrP^C–PrP^Sc interactions, which then leads to the misfolded isoform. We have therefore examined the molecular motions of mouse PrP^C, residues 113–231, in solution, using ¹⁵N NMR relaxation measurements. A truncated fragment has been used to eliminate the effect of the 90‐residue unstructured tail of PrP^C so the dynamics of the structured domain can be studied in isolation. ¹⁵N longitudinal (T₁) and transverse relaxation (T₂) times as well as the proton‐nitrogen nuclear Overhauser effects have been used to calculate the spectral density at three frequencies, 0, ω_N, and 0.87ω_H. Spectral densities at each residue indicate various time‐scale motions of the main‐chain. Even within the structured domain of PrP^C, a diverse range of motions are observed. We find that removal of the tail increases T₂ relaxation times significantly indicating that the tail is responsible for shortening of T₂ times in full‐length PrP^C. The truncated fragment of PrP has facilitated the determination of meaningful order parameters (S²) from the relaxation data and shows for the first time that all three helices in PrP^C have similar rigidity. Slow conformational fluctuations of mouse PrP^C are localized to a distinct region that involves residues 171 and 172. Interestingly, residues 170–175 have been identified as a segment within PrP that will form a steric zipper, believed to be the fundamental amyloid unit. The flexibility within these residues could facilitate the PrP^C–PrP^Sc recognition process during fibril elongation.