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NMR characterization of the full‐length recombinant murine prion protein, m PrP(23–231)
Author(s) -
Riek Roland,
Hornemann Simone,
Wider Gerhard,
Glockshuber Rudi,
Wüthrich Kurt
Publication year - 1997
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/s0014-5793(97)00920-4
Subject(s) - recombinant dna , prion protein , chemistry , microbiology and biotechnology , virology , biology , biochemistry , medicine , pathology , gene , disease
The recombinant murine prion protein, m PrP(23–231), was expressed in E. coli with uniform 15 N‐labeling. NMR experiments showed that the previously determined globular three‐dimensional structure of the C‐terminal domain m PrP(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 m PrP(23–231). Coincidence of corresponding sequential and medium‐range nuclear Overhauser effects (NOE) showed that the helical secondary structures previously identified in m PrP(121–231) are also present in m PrP(23–231), and near‐identity of corresponding amide nitrogen and amide proton chemical shifts indicates that the three‐dimensional fold of m PrP(121–231) is also preserved in the intact protein. The linewidths in heteronuclear 1 H– 15 N correlation spectra and 15 N{ 1 H}‐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 m PrP(23–231) outside of this domain.