Solution structure of Syrian hamster prion protein rPrP(90-231)

NMR has been used to refine the structure of Syrian hamster (SHa) prion pro tein rPrP(90-231), which is commensurate with the infectious protease-resis tant core of the scrapie prion protein PrPSc. The structure of rPrP(90-231) , refolded to resemble the normal cellular isoform PrPC spectroscopically a nd immunologically, has been studied using multidimensional NMR; initial re sults were published [James et al. (1997) Proc. Natl. Acad. Sci. U.S.A. 94, 10086-10091]. We now report refinement with better definition revealing im portant structural and dynamic features which can be related to biological observations pertinent to prion diseases. Structure refinement was based on 2778 unambiguously assigned nuclear Overhauser effect (NOE) connectivities , 297 ambiguous NOE restraints, and 63 scalar coupling constants ((3)J(HNHa )). The structure is represented by an ensemble of 25 best-scoring structur es from 100 structures calculated using ARIA/X-PLOR and further refined wit h restrained molecular dynamics using the AMBER 4.1 force field with an exp licit shell of water molecules. The rPrP(90-231) structure features a core domain (residues 125-228), with a backbone atomic root-mean-square deviatio n (RMSD) of 0.67 Angstrom, consisting of three alpha-helices (residues 144- 154, 172-193, and 200-227) and two short antiparallel beta-strands (residue s 129-131 and 161-163). The N-terminus (residues 90-119) is largely unstruc tured despite some sparse and weak medium-range NOEs implying the existence of bends or turns. The transition region between the core domain and flexi ble N-terminus, i.e., residues 113-128, consists of hydrophobic residues or glycines and does not adopt any regular secondary structure in aqueous sol ution. There are about 30 medium- and long-range NOEs within this hydrophob ic cluster, so it clearly manifests structure. Multiple discrete conformati ons are evident, implying the possible existence of one or more metastable states, which may feature in conversion of PrPC to PrPSc. To obtain a more comprehensive picture of rPrP(90-231), dynamics have been studied using ami de hydrogen-deuterium exchange and N-15 NMR relaxation times (T-1 and T-2) and N-15{H-1} NOE measurements. Comparison of the structure with previous r eports suggests sequence-dependent features that may be reflected in a spec ies barrier to prion disease transmission.