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Partial 13 C and 15 N Chemical‐Shift Assignments of the Disulfide‐Bond‐Forming Enzyme DsbB by 3D Magic‐Angle Spinning NMR Spectroscopy
Author(s) -
Li Ying,
Berthold Deborah A.,
Frericks Heather L.,
Gennis Robert B.,
Rienstra Chad M.
Publication year - 2007
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.200600484
Subject(s) - chemistry , magic angle spinning , crystallography , solid state nuclear magnetic resonance , chemical shift , nuclear magnetic resonance spectroscopy , nmr spectra database , periplasmic space , stereochemistry , escherichia coli , spectral line , nuclear magnetic resonance , physics , biochemistry , astronomy , gene
DsbB is a 20 kDa Escherichia coli inner‐membrane protein that catalyzes disulfide‐bond formation in periplasmic proteins. We report highly resolved, multidimensional magic‐angle spinning NMR spectra at 750 MHz 1 H frequency, which enable partial 13 C and 15 N chemical‐shift assignments of the signals. The narrow line widths observed indicate excellent microscopic order of the protein sample, suitable for full structure determination by solid‐state NMR. Experiments were performed exclusively on uniformly 13 C, 15 N‐labeled DsbB. Chemical‐shift‐correlation experiments based on dipolar transfer yielded strong signals in the 3D spectra, many of which have been site‐specifically assigned to the four transmembrane helices of DsbB. Significant numbers of additional residues have been assigned to stretches of amino acids, although not yet placed in the amino acid sequence. We also report the temperature dependence of signal intensities from −50 °C to 0 °C, a range over which samples of DsbB are highly stable. Structural and dynamic information derived from SSNMR studies can give insight into DsbB in a state that so far has not been successfully crystallized.