NMR Characterization of the Full-Length Recombinant Murine Prion Protein, mPrP(23-231)

FEBS Lett. 1997 Aug 18;413(2):282-8. doi: 10.1016/s0014-5793(97)00920-4.

Abstract

The recombinant murine prion protein, mPrP(23-231), was expressed in E. coli with uniform 15N-labeling. NMR experiments showed that the previously determined globular three-dimensional structure of the C-terminal domain mPrP(121-231) is preserved in the intact protein, and that the N-terminal polypeptide segment 23-120 is flexibly disordered. This structural information is based on nearly complete sequence-specific assignments for the backbone amide nitrogens, amide protons and alpha-protols of the polypeptide segment of residues 121-231 in mPrP(23-231). Coincidence of corresponding sequential and medium-range nuclear Overhauser effects (NOE) showed that the helical secondary structures previously identified in mPrP(121-231) are also present in mPrP(23-231), and near-identity of corresponding amide nitrogen and amide proton chemical shifts indicates that the three-dimensional fold of mPrP(121-231) is also preserved in the intact protein. The linewidths in heteronuclear 1H-15N correlation spectra and 15N[1H]-NOEs showed that the well structured residues 126-230 have correlation times of several nanoseconds, as is typical for small globular proteins, whereas correlation times shorter than 1 nanosecond were observed for all residues of mPrP(23-231) outside of this domain.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Magnetic Resonance Spectroscopy / methods*
  • Mice
  • Models, Molecular
  • Molecular Sequence Data
  • PrPC Proteins / chemistry*
  • Protein Conformation
  • Protein Structure, Secondary*
  • Recombinant Fusion Proteins / chemistry

Substances

  • PrPC Proteins
  • Recombinant Fusion Proteins