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51.
Evgeniy S. Salnikov Olivier Ouari Eline Koers Hiba Sarrouj Trent Franks Melanie Rosay Shane Pawsey Christian Reiter Priyanga Bandara Hartmut Oschkinat Paul Tordo Frank Engelke Burkhard Bechinger 《Applied magnetic resonance》2012,43(1-2):91-106
Dynamic nuclear polarization (DNP)/solid-state nuclear magnetic resonance (NMR) spectroscopy bears great potential for the investigation of membrane-associated polypeptides which can often be produced only in small amounts and which need to be ‘diluted’ in lipid bilayer environments to adopt or maintain their functional structure. Here we present investigations using biradicals, such as TOTAPOL and bTbK, for solid-state NMR signal enhancement using DNP in the context of lipid membranes. By transferring polarization from electron to nuclear spins using microwave irradiation signal enhancement factors of up to 13 are obtained with TOTAPOL and up to 17 with bTbK. The possible reasons why these factors are below those obtained in glassy samples of bulk solvents (40–60 under similar conditions) are evaluated and discussed. In order to further ameliorate the enhancement factors the physico-chemical characteristics of TEMPOL, TOTAPOL, bTbK, and bCTbK, such as their partitioning between hydrophilic and hydrophobic solvents or their stability under different environmental conditions are presented. Finally, having provided proof-of-concept that DNP/solid-state NMR measurements can be performed with oriented membrane samples work in progress is presented on the development of a flat-coil probe for DNP/solid-state NMR experiments on oriented membranes. 相似文献
52.
Franks WT Wylie BJ Stellfox SA Rienstra CM 《Journal of the American Chemical Society》2006,128(10):3154-3155
Structural studies of uniformly labeled proteins by magic-angle spinning NMR spectroscopy have rapidly matured in recent years. Site-specific chemical shifts of several proteins have been assigned and structures determined from 2D or 3D data sets containing internuclear distance information. Here we demonstrate the application of a complementary technique for constraining protein backbone geometry using a site-resolved 3D dipolar-shift pulse sequence. The dipolar line shapes report on the relative orientations of 1H-15N[i] to 1H-15N[i+1] dipole vectors, constraining the torsion angles phi[i] and psi[i]. In addition, from the same 3D data set, several 1H-15N[i] to1H-15N[i+2] line shapes are extracted to constrain the torsion angles phi[i], psi[i], phi[i+1], and psi[i+1]. We report results for the majority of sites in the 56-residue beta1 immunoglobulin binding domain of protein G (GB1), using 3D experiments at 600 MHz 1H frequency. Excellent agreement between the SSNMR results and a new 1.14 A crystal structure illustrate the general potential of this technique for high-resolution structural refinement of solid proteins. 相似文献
53.
Amide 15N chemical shift anisotropy (CSA) tensors provide quantitative insight into protein structure and dynamics. Experimental determinations of 15N CSA tensors in biologically relevant molecules have typically been performed by NMR relaxation studies in solution, goniometric analysis of single-crystal spectra, or slow magic-angle spinning (MAS) NMR experiments of microcrystalline samples. Here we present measurements of 15N CSA tensor magnitudes in a protein of known structure by three-dimensional MAS solid-state NMR. Isotropic 15N, 13C alpha, and 13C' chemical shifts in two dimensions resolve site-specific backbone amide recoupled CSA line shapes in the third dimension. Application of the experiments to the 56-residue beta1 immunoglobulin binding domain of protein G (GB1) enabled 91 independent determinations of 15N tensors at 51 of the 55 backbone amide sites, for which 15N-13C alpha and/or 15N-13C' cross-peaks were resolved in the two-dimensional experiment. For 37 15N signals, both intra- and interresidue correlations were resolved, enabling direct comparison of two experimental data sets to enhance measurement precision. Systematic variations between beta-sheet and alpha-helix residues are observed; the average value for the anisotropy parameter, delta (delta = delta(zz) - delta(iso)), for alpha-helical residues is 6 ppm greater than that for the beta-sheet residues. The results show a variation in delta of 15N amide backbone sites between -77 and -115 ppm, with an average value of -103.5 ppm. Some sites (e.g., G41) display smaller anisotropy due to backbone dynamics. In contrast, we observe an unusually large 15N tensor for K50, a residue that has an atypical, positive value for the backbone phi torsion angle. To our knowledge, this is the most complete experimental analysis of 15N CSA magnitude to date in a solid protein. The availability of previous high-resolution crystal and solution NMR structures, as well as detailed solid-state NMR studies, will enhance the value of these measurements as a benchmark for the development of ab initio calculations of amide 15N shielding tensor magnitudes. 相似文献
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This study addresses the possibility that ice-Ic may form spontaneously and directly during the cooling of previously freeze-concentrated aqueous solutions, at temperatures where the residual unfrozen water is likely to exist in small, isolated domains. It is shown that on cooling a dilute aqueous glucose solution, ice-Ih crystallises initially, but when the solution has been freeze-concentrated to ca.10 mol %, further cooling results in the slow nucleation and growth of ice-Ic in the residual water domains, rather than on the pre-existing ice-Ih crystal surfaces. 相似文献
57.
58.
Murphy WJ Franks JR Berger EH Behar A Casali JG Dixon-Ernst C Krieg EF Mozo BT Royster JD Royster LH Simon SD Stephenson C 《The Journal of the Acoustical Society of America》2004,115(1):311-323
The mandate of ASA Working Group S12/WG11 has been to develop "laboratory and/or field procedure(s) that yield useful estimates of field performance" of hearing protection devices (HPDs). A real-ear attenuation at threshold procedure was selected, devised, tested for one earmuff and three earplugs via an interlaboratory study involving five laboratories and 147 subjects, and incorporated into a new standard that was approved in 1997 [Royster et al., "Development of a new standard laboratory protocol for estimating the field attenuation of hearing protection devices. Part I. Research of Working Group 11, Accredited Standards Committee S 12, Noise," J. Acoust. Soc. Am. 99, 1506-1526; ANSI, S12.6-1997, "American National Standard method for measuring real-ear attenuation of hearing protectors" (American National Standards Institute, New York, 1997)]. The subject-fit methodology of ANSI S12.6-1997 relies upon listeners who are audiometrically proficient, but inexperienced in the use of HPDs. Whenever a new method is adopted, it is important to know the effects of variability on the power of the measurements. In evaluation of protector noise reduction determined by experimenter-fit, informed-user-fit, and subject-fit methods, interlaboratory reproducibility was found to be best for the subject-fit method. Formulas were derived for determining the minimum detectable difference between attenuation measurements and for determining the number of subjects necessary to achieve a selected level of precision. For a precision of 6 dB, the study found that the minimum number of subjects was 4 for the Bilsom UF-1 earmuff, 10 for the E.A.R Classic earplug, 31 for the Willson EP100 earplug, and 22 for the PlasMed V-51R earplug. 相似文献
59.
60.
Nicholas NJ Franks GV Ducker WA 《Langmuir : the ACS journal of surfaces and colloids》2012,28(18):7189-7196
We examine the hypothesis that selective adsorption to a particular face of ZnO is responsible for the ability of small organic molecules to control the aspect ratio of ZnO crystals during hydrothermal synthesis. Large, single crystals of ZnO were prepared such that the vast majority of a surface consisted of a single crystal plane, as shown by atomic force microscopy, and the adsorption to a single crystal plane was determined by attenuated total reflectance spectroscopy. The results show that citrate strongly and selectively adsorbs to the (0001) face. Similarly, results show that ethylenediamine selectively adsorbs to the (1010) face. Each of these results separately shows a correlation between selective adsorption to and growth of large areas of a particular face, and thus, each result is consistent with the proposed hypothesis. 相似文献