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Methionine sulfoxide reductases (Msr) reduce methionine sulfoxide (MetSO)-containing proteins, back to methionine (Met). MsrAs are stereospecific for the S epimer whereas MsrBs reduce the R epimer of MetSO. Although structurally unrelated, the Msrs characterized so far display a similar catalytic mechanism with formation of a sulfenic intermediate on the catalytic cysteine and a concomitant release of Met, followed by formation of at least one intramolecular disulfide bond (between the catalytic and a recycling cysteine), which is then reduced by thioredoxin. In the case of the MsrA from Escherichia coli, two disulfide bonds are formed, i.e. first between the catalytic Cys51 and the recycling Cys198 and then between Cys198 and the second recycling Cys206. Three crystal structures including E. coli and Mycobacterium tuberculosis MsrAs, which, for the latter, possesses only the unique recycling Cys198, have been solved so far. In these structures, the distances between the cysteine residues involved in the catalytic mechanism are too large to allow formation of the intramolecular disulfide bonds. Here structural and dynamical NMR studies of the reduced wild-type and the oxidized (Cys51-Cys198) forms of C86S/C206S MsrA from E. coli have been carried out. The mapping of MetSO substrate-bound C51A MsrA has also been performed. The data support (1) a conformational switch occurring subsequently to sulfenic acid formation and/or Met release that would be a prerequisite to form the Cys51-Cys198 bond and, (2) a high mobility of the C-terminal part of the Cys51-Cys198 oxidized form that would favor formation of the second Cys198-Cys206 disulfide bond.

A gradient-based sequence is proposed for efficiently filtering out all quantities except the longitudinal two-spin order, created by csa/dipolar cross-correlation rates. The dipolar interaction is between a proton and a heteronucleus, the csa being generally the one of the heteronucleus. Proton detection is carried out to benefit from maximum sensitivity. The resulting two-dimensional spectrum has the same aspect as an HMBC spectrum, implying the existence of a J coupling between the two considered nuclei, but here cross-peaks indicate the strength of the relevant csa/dipolar cross-correlation rate. The method is especially interesting in the case of medium-sized molecules where a given heteronucleus is subjected to several csa/dipolar cross-correlation effects, along with the corresponding J couplings.

This paper presents a non-iterative, fast, and almost automated time-data analysis method for NMR spectroscopy, based on a new adaptive implementation of high resolution methods used in spectral subbands. It is intended to avoid the choice of the decimation factor (or the width of the spectral windows) which, in the case of a uniform decomposition, strongly conditions the estimation results, and to diminish the computational burden. It is achieved through successive decimation/estimation stages each followed by a test procedure in order to decide whether or not the process should continue. The proposed test is based on a local spectral flatness measure of the estimation residuals. This stop-criterion involves an a posteriori validation of the estimation, thus the method proposed allows one to obtain a better detection rate at a lower complexity comparatively to other stopping rules, while preserving a reasonable estimation variance. Moreover, the reliability of the fitting algorithms considered is improved, by decreasing the influence of the model order and the number of false detections. Finally, the method is more efficient than Fourier transform (FT) at low signal-to-noise ratio (SNR). The effectiveness of the method is demonstrated by analyzing a simulation signal and raw carbon-13 experimental data.

The phase-twisted lines, resulting from a double complex Fourier transform, can greatly alter the quality of two-dimensional spectra. This problem, which is generally overcome by experimental procedures (requiring twice as much time as a standard measurement), is inevitable in the two-dimensional J-resolved experiment. This experiment is revisited theoretically and a postprocessing treatment, leading to pure 2D J-spectra, is deduced. Quantitative fully J-decoupled homonuclear spectra are accordingly obtained by means of a projection onto the F2 axis after a 45 degrees tilt., (Copyright 1999 Academic Press.)

It is shown that fully J-decoupled homonuclear spectra involving Lorentzian lines can be readily obtained by straightforward processing of the 2D data arising from a conventional spin echo sequence (pi/2-t(1)/2-pi-t(1)/2-Acq(t(2))) used in the so-called J-resolved experiment. The method simply rests on power spectra with the drawback of lines having meaningless relative intensities. In principle, the experiment should also yield transverse relaxation times. Several tests demonstrate that this is not so, due to pulse imperfections and nonresolved long-range J couplings. Conversely, longitudinal and rotating frame relaxation times can be easily determined by means of an appropriate preparation period (for instance, a saturation-recovery period in the case of longitudinal relaxation) inserted before the 2D spin echo sequence. Since one is dealing with a single line per nucleus, relaxation measurements become reliable and accurate., (Copyright 1999 Academic Press.)

It is shown that the proton chemical-shift anisotropy of hydrate crystals affect dipolar powder pattern at 7.0 T. This may be clearly observed via an asymmetric envelope of dipolar spinning sidebands in magic-angle spinning proton spectra.