NMR characterization of the full-length recombinant murine prion protein, mPrP(23–231)
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 α-protons 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:
-
FEBS Letters
- Pub Date:
- January 1997
- DOI:
- 10.1016/S0014-5793(97)00920-4
- Bibcode:
- 1997FEBSL.413..282R
- Keywords:
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- Prion protein;
- NMR;
- Protein structure and dynamics;
- Transmissible spongiform encephalophaties;
- Correlation time