A large-sample-volume constant-flow magic angle sample spinning (CF-MAS) NMR probe is reported for in situ studies of the reaction dynamics, stable intermediates/transition states, and mechanisms of catalytic reactions. In our approach, the reactants are introduced into the catalyst bed using a fixed tube at one end of the MAS rotor while a second fixed tube, linked to a vacuum pump, is attached at the other end of the rotor. The pressure difference between both ends of the catalyst bed inside the sample cell space forces the reactants flowing through the catalyst bed, which improves the diffusion of the reactants and products. This design allows the use of a large sample volume for enhanced sensitivity and thus permitting in situ C-13 CF-MAS studies at natural abundance. As an example of application, we show that reactants, products and reaction transition states associated with the 2-butanol dehydration reaction over a mesoporous silicalite supported heteropoly acid catalyst (HPA/meso-silicalite-1) can all be detected in a single C-13 CF-MAS NMR spectrum at natural abundance. Coke products can also be detected at natural C-13 abundance and under the stopped flow condition. Furthermore, H-1 CF-MAS NMR is used to identify the surface functional groups of HPA/meso-silicalite-1 under the condition of in situ drying. We also show that the reaction dynamics of 2-butanol dehydration using HPA/meso-silicalite-1 as a catalyst can be explored using H-1 CF-MAS NMR.

Methyl groups are thought to dominate the dynamics of proteins after slow collective modes of motion freeze out in a glass-transition process. In this work we investigate methyl group dynamics of a key hydrophobic core leucine residue in chicken villin headpiece subdomain protein at 140-4 K using deuteron NMR longitudinal relaxation measurements. A distinct increase in the apparent activation energy is observed at approximately 95 K, indicating an abrupt freezing of methyl group dynamics. Relaxation times at temperatures below 60 K are dominated by the deuteron tunneling mechanism.

Quantitative dynamics of methyl groups in 9-fluorenylmethyloxycarbonyl-leucine (FMOC-leu) have been analyzed and compared with earlier studies of methyl dynamics in chicken villin headpiece subdomain protein (HP36) labeled at L69, a key hydrophobic core position. A combination of deuteron solid-state nuclear magnetic resonance experiments over the temperature range of 7-324 K and computational modeling indicated that while the two compounds show the same modes of motions, there are marked differences in the best-fit parameters of these motions. One of the main results is that the crossover observed in the dynamics of the methyl groups in the HP36 sample at 170 K is absent in FMOC-leu. A second crossover at around 95-88 K is present in both samples. The differences in the behavior of the two compounds suggest that some of the features of methyl dynamics reflect the complexity of the protein hydrophobic core and are not determined solely by local interactions.

Advances in understanding of sensor and other modern complex materials are often enabled by new research tools. This paper highlights three capability development themes used to identify new research tools to be provided to users of the U. S. Department of Energy's Environmental Molecular Sciences Laboratory. These capability development directions address the importance of dynamic measurements in realistic environments, the need for increased resolution in three dimensional analyses as well as the importance of linking theory and experiment. Capability development involves expanding the range of operation for a number of important techniques, developing and applying new capabilities, and advancing methods of data processing. Examples of current developments are provided including those related to magnetic resonance, x-ray diffraction, application of a focused beam capability to fuel cell aging, and near real time analysis of x-ray photoelectron spectroscopy spectra.

With the goal of investigating dynamical features of hydrophobic cores of proteins over a wide range of temperatures, the chicken villin headpiece subdomain protein (HP36) was labeled at a "single" site corresponding to any one of the two C(delta)D(3) groups of leucine-69, which is located in a key position of the core. The main techniques employed are deuteron NMR quadrupolar echo line shape analysis, and T(1Z) (Zeeman) and T(1Q) (quadrupolar order) relaxation experiments performed at 11.7 and 17.6 T over the temperature range of 112 to 298 K. The experimental data are compared with computer simulations. The deuteron line shapes give an excellent fit to a three-mode motional model that consists of (a) fast three-site rotational jumps about the pseudo C(3) methyl spinning axis, (b) slower reorientation of the spinning axis, described by diffusion along a restricted arc, and (c) large angle jumps between traces of rotameric conformers. Relaxation behavior is described by a phenomenological distribution of activation energies for three-site hops at high temperatures that collapses to a single, distinctly smaller value for lower temperatures.

Motivated by a recent proposal by Sushkov and co-workers [Phys. Rev. A 72, 034501 (2005); 73, 022107 (2006)] to search for a P,T-violating Schiff moment of the Pb-207 nucleus in a ferroelectric solid, we have carried out a high-field nuclear magnetic resonance study of the longitudinal and transverse spin relaxation of the lead nuclei from room temperature down to 10 K for powder samples of lead titanate (PT), lead zirconium titanate, and a PT monocrystal. For all powder samples and independently of temperature, transverse relaxation times were found to be T-2 approximate to 1.5 ms, while the longitudinal relaxation times exhibited a temperature dependence, with T-1 of over an hour at the lowest temperatures, decreasing to T-1 approximate to 7 s at room temperature. At high temperatures, the observed behavior is consistent with a two-phonon Raman process, while in the low-temperature limit, the relaxation appears to be dominated by a single-phonon (direct) process involving magnetic impurities. We discuss the implications of the results for the Schiff-moment search.

Large macromolecular machines are among the most important and challenging targets for structural and mechanistic analyses. Consequently, there is great interest in development of NMR methods for the study of multicomponent systems in the 50-500 kDa range. Biochemical methods also must be developed in concert to produce such systems in selectively labeled form. Here, we present (1)H/(13)C-HSQC spectra of protonated methyl groups in a model system that mimics molecular weights up to approximately 560 kDa. Signals from side chain methyl groups of Ile, Leu, and Val residues are clearly detectable at correlation times up to approximately 330 ns. We have also developed a biochemical procedure to produce the 240 kDa, heteroheptameric Arp2/3 actin nucleation complex selectively labeled at one subunit and obtained (1)H/(13)C-HSQC spectra of this assembly. Sensitivity in spectra of both the Arp2/3 complex and the model system indicate that methyl groups will be useful sources of information in nonsymmetric systems with molecular weights greater than 600 kDa at concentrations less than 100 microM. Methyl analyses will complement TROSY and CRINEPT analyses of amides in NMR studies of structure and molecular interactions of extremely large macromolecules and assemblies.

Using a Taylor series expansion of the phase shift of a moving isochromatic spin group in the presence of a magnetic field gradient, the refocusing effects of the Carr‐Purcell‐Meiboom‐Gill pulse sequence on stationary nuclei and those with constant, rectilinear velocity are readily demonstrated. Continuing the analysis to higher orders of motion reveals that nuclei with a constant, rectilinear acceleration have a phase shift at the spin echoes which increases linearly with echo number. Constant, rectilinear jerk (the time rate of change of acceleration) leads to an increase in phase shift from echo to echo which is quadratic in nature with an overlying reduction of the odd‐numbered echoes by a constant amount. Motion parameters may be measured by parameter identification techniques. These principles may be applied to phase‐sensitive NMR imaging.© 1985 Academic Press,Inc.

We present a method for producing accurate calculated T1 and spin density nuclear magnetic resonance images. A modified Carr-Purcell-Meiboom-Gill pulse sequence is used to obtain a series of images containing both T1 and T2 dependence. The image series is first analyzed to remove the T2 dependence. The resulting images are then analyzed, pixel by pixel, to generate an image containing T1 values and an image containing values proportional to spin density (SD). Tests performed on two phantoms containing solutions of various known T1's and H2O/D2O concentrations indicate that the T1 image values are accurate to better than 11% and the relative SD values agree to within one standard deviation.

A prospective clinical study was performed in 32 patients with multiple sclerosis (MS) to evaluate the sensitivity of lesion detection and accuracy of lesion localization by neurologic examination, delayed enhanced computed tomography (CT) with a double dose of contrast material, and proton magnetic resonance (MR) imaging. After neurologic examination patients were classified by probability of MS (possible, four patients; probable, three patients; and definite, 25 patients) and by disease activity (acute, chronic with acute exacerbation, or chronic progressive). Subsequently they underwent delayed enhanced CT scanning and MR imaging with more than one spin-echo technique. In five of seven patients with possible or probable MS, both MR imaging and delayed enhanced CT were negative. In 25 cases of definite MS, MR imaging detected pathology in 19 (76%) cases, while CT detected lesions in 15 (60%) of 25 cases. In acute lesions (acute or chronic with acute exacerbation), the two techniques were of similar sensitivity (delayed CT was positive in 65% and MR imaging in 60%), while in chronic progressive MS, MR imaging was superior in lesion detection (MR imaging positive in 75%; delayed CT in 25%). While most lesions (55%) were seen in corresponding locations in both studies, neither MR nor delayed CT correlated well with lesion localization by neurologic examination because a large number of asymptomatic lesions were imaged and many symptomatic lesions were undetected.(ABSTRACT TRUNCATED AT 250 WORDS)

Magnetic Resonance Imaging (MRI) affords a considerable improvement in image contrast over other methods by virtue of the intrinsic NMR parameters spin density, T1, and T2. However, the clinical utility of routine quantification of these parameters is currently unknown. Calculated T2 images might afford additional disease specific information provided the calculation algorithm generates accurate T2 values. In this study, calculated T2 images of a MnCl2 phantom (spanning a T2 range of interest of 45.7 ms to 346.6 ms at 6 MHz) were generated utilizing a variety of calculation algorithms based upon a data set of 32 sequential spin-echo (SE) images. In general, when utilizing only the earliest sequential SE after the 90 degree pulse for the T2 calculation, the greater the number of SE used in the calculation algorithm, regardless of how they were averaged, the more accurate and less noisy was the calculated image. When only limited numbers of SE were used in the calculation algorithm, accuracy and noise varied with the choice of TE suggesting that there may be optimal timings for TE for a particular T2 range of interest. Forty-two calculated T2 head images of normal subjects, based upon data sets of 16 sequential SE, were evaluated for the T2 values of normal brain. These were compared to T2 images calculated via 7 different algorithms based upon 16 SE data sets from two patients with CNS pathology. An optimal algorithm was identified in which 16 SE Carr-Purcell-Meiboom-Gill (CPMG) were averaged into two images for the T2 calculation. With this algorithm, calculated images could be generated efficiently which were accurate and relatively noise free. The availability of such images maximized whatever disease specificity, and thus clinical utility, T2 information affords.

AbstractThe synthesis of 1, 4, 9‐triazaphenoxathiin and the analysis of the two‐dimensional J‐resolved (2DJ) 1H‐nmr spectra of the molecule at 100 and 200 MHz are reported. Due to the strong coupling of the ABX and AB spin systems, additional resonances are observed in the 100 MHz 2DJ spectrum which are diminished in intensity and shifted further from F1 = O Hz in the case of the ABX system at 200 MHz, or which have disappeared in the 200 MHz spectrum in the case of the AB spin system. Calculated frequencies and intensities of the resonances due to strong coupling were found to be in excellent agreement with the observed data.

The solution structure of rifaximin and its derivative rifaximin OR (open ring) was determined by combining NMR experimental results, theoretical simulation of two-dimensional NMR spectra by complete relaxation matrix analysis (corma), and molecular dynamics calculations.In this study the structural rearrangements due to the opening of the aliphatic chain of rifaximin after the reduction process to form rifaximin OR were investigated.Close spatial proximity of CH3(14) and H28b protons detected by 2D-ROESY spectrum of rifaximin OR, which was not present in rifaximin and the down-field shift of CH3(34) protons in rifaximin OR 1H spectrum were crucial to understand the structural modifications, which occurred within the system. The aliphatic chain of rifaximin OR was found to be no longer symmetrical with respect to the aromatic moiety. Although no dramatic structural rearrangements were detected, the aliphatic chain moved toward CH3(14), causing a reduction of the aromatic shielding contribution in particular on CH3(34).