共查询到20条相似文献,搜索用时 15 毫秒
1.
Günter Hempel Matthias Wobst Gunter Israel Horst Schneider 《Macromolecular Symposia》1993,72(1):161-189
Two polysiloxanes with mesogenic side chains were investigated with polarization microscopy, DSC, 1H and 13C NMR. From proton NMR the global order parameter of one sample (the other does not orient itself in the magnetic field of 2.1 T) and some information about phase transitions was obtained. 13C NMR yielded selective order parameters with respect to different atomic positions. The order parameter varies slightly also within the mesogenic unit, but much stronger within the spacer (odd-even effect). With the help of T1ρ experiments some molecular motions had been detected and characterized (main-chain motion, over-head rotation of the mesogenic unit, methyl rotation, side-chain rotation). 相似文献
2.
New NMR experiments for the measurement of side-chain dynamics in high molecular weight ( approximately 100 kDa) proteins are presented. The experiments quantify (2)H spin relaxation rates in (13)CH(2)D or (13)CHD(2) methyl isotopomers and, for applications to large systems, offer significant gains both in sensitivity (2-3-fold) and resolution over previously published HSQC schemes. The methodology has been applied to investigate Ile dynamics in the 723-residue, single polypeptide chain enzyme, malate synthase G. Methyl-axis order parameters, S(axis), characterizing the amplitudes of motion of the methyl groups, have been derived from both (13)CH(2)D and (13)CHD(2) probes and are in excellent agreement. The distribution of order parameters is trimodal, reflecting the range of dynamics that are available to Ile residues. A reasonable correlation is noted between and inverse temperature factors from X-ray studies of the enzyme. The proposed methodology significantly extends the range of protein systems for which side-chain dynamics can be studied. 相似文献
3.
We studied the interaction of mono- and polyunsaturated phosphatidylcholines with rhodopsin by 1H NMR saturation transfer difference spectroscopy with magic angle spinning (STD-MAS NMR). The results indicate a strong preference for interaction of rhodopsin with the polyunsaturated docosahexaenoic acid. 相似文献
4.
Hansen AL Lundström P Velyvis A Kay LE 《Journal of the American Chemical Society》2012,134(6):3178-3189
A Carr-Purcell-Meiboom-Gill relaxation dispersion experiment is presented for quantifying millisecond time-scale chemical exchange at side-chain (1)H positions in proteins. Such experiments are not possible in a fully protonated molecule because of magnetization evolution from homonuclear scalar couplings that interferes with the extraction of accurate transverse relaxation rates. It is shown, however, that by using a labeling strategy whereby proteins are produced using {(13)C,(1)H}-glucose and D(2)O a significant number of 'isolated' side-chain (1)H spins are generated, eliminating such effects. It thus becomes possible to record (1)H dispersion profiles at the β positions of Asx, Cys, Ser, His, Phe, Tyr, and Trp as well as the γ positions of Glx, in addition to the methyl side-chain moieties. This brings the total of amino acid side-chain positions that can be simultaneously probed using a single (1)H dispersion experiment to 16. The utility of the approach is demonstrated with an application to the four-helix bundle colicin E7 immunity protein, Im7, which folds via a partially structured low populated intermediate that interconverts with the folded, ground state on the millisecond time-scale. The extracted (1)H chemical shift differences at side-chain positions provide valuable restraints in structural studies of invisible, excited states, complementing backbone chemical shifts that are available from existing relaxation dispersion experiments. 相似文献
5.
Reif B Xue Y Agarwal V Pavlova MS Hologne M Diehl A Ryabov YE Skrynnikov NR 《Journal of the American Chemical Society》2006,128(38):12354-12355
Rapid advances in solid-state MAS NMR made it possible to probe protein dynamics on a per-residue basis, similar to solution experiments. In this work we compare methyl 2H relaxation rates measured in the solid and liquid samples of alpha-spectrin SH3 domain. The solution data are treated using a model-free approach to separate the contributions from the overall molecular tumbling and fast internal motion. The latter part forms the basis for comparison with the solid-state data. Although the accuracy of solid-state measurements is limited by deuterium spin diffusion, the results suggest a significant similarity between methyl dynamics in the two samples. This is a potentially important observation, preparing the ground for combined analysis of the dynamics data by solid- and solution-state NMR. 相似文献
6.
7.
By using a combination of 1H NMR spectroscopy, two-dimensional heteronuclear single-quantum coherence-resolved (1)H{(13)C} and homonuclear rotating-frame Overhauser enhancement NMR correlation experiments with diffusion ordered spectroscopy (DOSY), the location and distribution of a hydrophobic drug, paeonol, have been established with respect to the methyl groups of the poly(ethylene oxide)-poly(propylene oxide) -poly(ethylene oxide) (PEO-PPO-PEO) triblock copolymer. The interaction between them is adjustable according to the different temperature-dependent hydrophilicities or hydrophobicities of the triblock copolymer components. On the other hand, such interactions influence the self-assembly properties of the block copolymer amphiphiles in solution. The amount of anhydrous methyl groups of PPO segments shows an increase with increasing paeonol concentration. It was also demonstrated that the shell-crosslinking of the Pluronic polymer has an effect in increasing the amount of anhydrous methyl groups and thus increasing the hydrophobicity of Pluronic micelles. This might be the deeper reason underlying the increase in drug-loading capacity and prolongation in release time of Pluronic micelles for drug delivery after the shell-crosslinking. Changes in self-diffusion coefficients of paeonol with varying copolymer concentrations and types were also determined by the diffusion-based NMR DOSY technique, and values of K(a), DeltaG, and n were calculated. 相似文献
8.
Michael Wübbenhorst Ernout M. Van Koten John C. Jansen Wim Mijs Jan van Turnhout 《Macromolecular rapid communications》1997,18(2):139-147
The molecular dynamics of amorphous and liquid-crystalline (LC) side-chain polycarbonates was studied by dielectric spectroscopy at frequencies from 10−2 to 106 Hz and at temperatures from −160 to 180°C. ‘Model’ compounds containing no mesogenic side-groups showed two relaxations, which originate from the carbonate group (α, βm-relaxation). By contrast, in LC-polycarbonates bearing a mesogenic nitrostilbene side group around and above the glass transition temperature Tg up to three relaxation modes were distinguished (α-, λ1-, λ2-process); below Tg four secondary relaxations (γ-, βm-, βs-, βsc-relaxation) were observed. The γ-relaxation was found only in compounds possessing an aliphatic spacer linked to the backbone by an ether bond. Apart from βm-, two additional β-processes were identified as relaxations associated with the mesogenic unit in the glassy (βs) or in the crystalline state (βsc). 相似文献
9.
An NMR experiment is presented for the measurement of the time scale of methyl side-chain dynamics in proteins that are labeled with methyl groups of the (13)CHD(2) variety. The measurement is accomplished by selecting a magnetization mode that to excellent approximation relaxes in a single-exponential manner with a T(1)-like rate. The combination of R(1)((13)CHD(2)) and R(2)((13)CHD(2)) (2)H relaxation rates facilitates the extraction of motional parameters from (13)CHD(2)-labeled proteins exclusively. The utility of the methodology is demonstrated with applications to proteins with tumbling times ranging from 2 ns (protein L, 7.5 kDa, 45 degrees C) to 54 ns (malate synthase G, 82 kDa, 37 degrees C); dynamics parameters are shown to be in excellent agreement with those obtained in (2)H NMR studies of other methyl isotopomers. A consistency relationship is found to exist between R(1)((13)CHD(2)) and the relaxation rates of pure longitudinal and quadrupolar order modes in (13)CH(2)D-labeled methyl groups, and experimental rates measured for a number of proteins are shown to be in excellent agreement with expectations based on theory. The present methodology extends the applicability of (2)H relaxation methods for the quantification of side-chain dynamics in high molecular weight proteins. 相似文献
10.
Dubois L Da Silva P Landon C Huber JG Ponchet M Vovelle F Berthault P Desvaux H 《Journal of the American Chemical Society》2004,126(48):15738-15746
The hydrophobic cavity of Lipid Transfer Protein 1 from Nicotiana tabacum is investigated in detail by NMR using xenon as a spy. The analysis of the (129)Xe chemical shifts and self-relaxation times gives evidence of protein-xenon interaction. Thermodynamics of the binding is characterized through the study of aliphatic (1)H and (13)C chemical shift variation as a function of xenon pressure. The binding constant is evaluated to 75.5 +/- 1.0 M(-1) at 293 K. The location of xenon inside the cavity is deduced from SPINOE experiments. The noble gas appears to occupy four sites, and xenon self-relaxation experiments indicate that it quickly jumps between different sites. The chemical shifts of amide protons and nitrogens also depend on the xenon concentration, either specifically or nonspecifically for atoms at the external surface of the protein. Yet, contrary to aliphatic atoms, they do not correspond to short-range interactions as confirmed by magnetization transfer experiments between laser-polarized xenon and protons in H(2)O. These (15)N chemical shift variations, used in combination with (15)N transverse self-relaxation rates to determine the lower limit of the binding rate, consequently reveal subtle changes in the structure of the protein upon binding. 相似文献
11.
A general framework is presented for the interpretation of NMR relaxation data of proteins. The method, termed isotropic reorientational eigenmode dynamics (iRED), relies on a principal component analysis of the isotropically averaged covariance matrix of the lattice functions of the spin interactions responsible for spin relaxation. The covariance matrix, which is evaluated using a molecular dynamics (MD) simulation, is diagonalized yielding reorientational eigenmodes and amplitudes that reveal detailed information about correlated protein dynamics. The eigenvalue distribution allows one to quantitatively assess whether overall and internal motions are statistically separable. To each eigenmode belongs a correlation time that can be adjusted to optimally reproduce experimental relaxation parameters. A key feature of the method is that it does not require separability of overall tumbling and internal motions, which makes it applicable to a wide range of systems, such as folded, partially folded, and unfolded biomolecular systems and other macromolecules in solution. The approach was applied to NMR relaxation data of ubiquitin collected at multiple magnetic fields in the native form and in the partially folded A-state using MD trajectories with lengths of 6 and 70 ns. The relaxation data of native ubiquitin are well reproduced after adjustment of the correlation times of the 10 largest eigenmodes. For this state, a high degree of separability between internal and overall motions is present as is reflected in large amplitude and collectivity gaps between internal and overall reorientational modes. In contrast, no such separability exists for the A-state. Residual overall tumbling motion involving the N-terminal beta-sheet and the central helix is observed for two of the largest modes only. By adjusting the correlation times of the 10 largest modes, a high degree of consistency between the experimental relaxation data and the iRED model is reached for this highly flexible biomolecule. 相似文献
12.
Rickard MA Pakoulev AV Mathew NA Kornau KM Wright JC 《The journal of physical chemistry. A》2007,111(7):1163-1166
Frequency-domain two-color triply vibrational enhanced four-wave mixing using a new phase-matching geometry discriminates against coherent multidimensional spectral features created solely by radiative transitions, spectrally resolves pathways with different numbers of coherence transfer steps, and temporally resolves modulations created by interference between coherence transfer pathways. Coherence transfer is a nonradiative transition where a superposition of quantum states evolves to a different superposition. The asymmetric and symmetric C[triple bond]O stretching modes of rhodium(I) dicarbonyl acetylacetonate are used as a model system for coherence transfer. A simplified theoretical model based on Redfield theory is used to describe the experimental results. 相似文献
13.
Julius Tsuwi Dietmar Appelhans Stefan Zschoche Rong-Chuan Zhuang Peter Friedel Liane Häußler Brigitte Voit Friedrich Kremer 《Colloid and polymer science》2005,283(12):1321-1333
A series of alternating maleimide (MI) copolymers with fluorinated side chains have been investigated using broadband dielectric
spectroscopy. The side chains consist of fluoroalkane (–C
x
F2x+1, x=1, 7, 9) end groups connected to the main chain via methylene spacers. The experiments were carried out in a frequency range
of 0.1 Hz to 10 MHz and at temperatures between 120 K and 500 K. The fluorinated MI copolymers show a fast sub-T
g (β) relaxation characterized by an Arrhenius-type temperature dependence with activation energy in the range of 30–37 kJ/mol.
Two more processes (α and δ-like) are observed, corresponding to independent relaxations of the main chain and the fluoroalkane
domains respectively. For shorter side chains, the δ-like process is not observed but instead another relaxation process,
α
S
, occurs at temperatures higher than either the α and δ-like processes. When compared with unfluorinated MI copolymers, the
fluorinated MI copolymers show the δ-like process and a slower β-relaxation unlike their unfluorinated counterparts. A model
to explain the molecular origin of the four processes is proposed, supplemented by differential scanning calorimetry and published
WAXS/SAXS data. 相似文献
14.
《Tetrahedron: Asymmetry》1999,10(15):2905-2912
This paper describes the conditions in which in situ NMR spectroscopy is a suitable technique to use when following the course of enzymatic transglycosylation reactions. Using this methodology, the reactions must be carried out in D2O. Our experiments indicate that the rate of the transglycosylation reaction is reduced in this solvent while the rate of the hydrolysis of the disaccharides produced is enhanced depending on the nature of the anomeric substituent. However, this undesirable effect is generally weak because the rates of the transglycosylation reactions are always faster than the rates of the hydrolysis whatever the solvent. The great advantage of NMR spectroscopy lies in its potential to detect, in a single experiment, all the components of the reaction without any disturbance of the reaction medium. 相似文献
15.
New relaxation dispersion experiments are presented that probe millisecond time-scale dynamical processes in proteins. The experiments measure the relaxation of (1)H-(15)N multiple-quantum coherence as a function of the rate of application of either (1)H or (15)N refocusing pulses during a constant time relaxation interval. In contrast to the dispersion profiles generated from more conventional (15)N((1)H) single-quantum relaxation experiments that depend on changes in (15)N((1)H) chemical shifts between exchanging states, (1)H-(15)N multiple-quantum dispersions are sensitive to changes in the chemical environments of both (1)H and (15)N spins. The resulting multiple-quantum relaxation dispersion profiles can, therefore, be quite different from those generated by single-quantum experiments, so that an analysis of both single- and multiple-quantum profiles together provides a powerful approach for obtaining robust measures of exchange parameters. This is particularly the case in applications to protonated proteins where other methods for studying exchange involving amide proton spins are negatively influenced by contributions from neighboring protons. The methodology is demonstrated on protonated and perdeuterated samples of a G48M mutant of the Fyn SH3 domain that exchanges between folded and unfolded states in solution. 相似文献
16.
Brath U Akke M Yang D Kay LE Mulder FA 《Journal of the American Chemical Society》2006,128(17):5718-5727
Transverse relaxation dispersion NMR spectroscopy can provide atom-specific information about time scales, populations, and the extent of structural reorganization in proteins under equilibrium conditions. A method is described that uses side-chain methyl groups as local reporters for conformational transitions taking place in the microsecond regime. The experiment measures carbon nuclear spin relaxation rates in the presence of continuous wave off-resonance irradiation, in proteins uniformly enriched with 13C, and partially randomly labeled with 2H. The method was applied to human FK-506 binding protein (FKBP12), which uses a common surface for binding substrates in its dual role as both an immunophilin and folding assistant. Conformational dynamics on a time scale of approximately 130 micros were detected for methyl groups located in the substrate binding pocket, demonstrating its plasticity in the absence of substrate. The spatial arrangement of affected side-chain atoms suggests that substrate recognition involves the rapid relative movement of the subdomain comprising residues Ala81-Thr96 and that the observed dynamics play an important role in facilitating the interaction of this protein with its many partners, including calcineurin. 相似文献
17.
Korzhnev DM Kloiber K Kanelis V Tugarinov V Kay LE 《Journal of the American Chemical Society》2004,126(12):3964-3973
A new CPMG-based multiple quantum relaxation dispersion experiment is presented for measuring millisecond dynamic processes at side-chain methyl positions in high molecular weight proteins. The experiment benefits from a methyl-TROSY effect in which cancellation of intramethyl dipole fields occurs, leading to methyl (13)C-(1)H correlation spectra of high sensitivity and resolution (Tugarinov, V.; Hwang, P. M.; Ollerenshaw, J. E.; Kay, L. E. J. Am. Chem. Soc. 2003, 125, 10420-10428). The utility of the methodology is illustrated with an application to a highly deuterated, methyl-protonated sample of malate synthase G, an 82 kDa enzyme consisting of a single polypeptide chain. A comparison of the sensitivity obtained using the present approach relative to existing HSQC-type (13)C single quantum dispersion experiments shows a gain of a factor of 5.4 on average, significantly increasing the range of applications for this methodology. 相似文献
18.
Joly AG Henyk M Beck KM Trevisanutto PE Sushko PV Hess WP Shluger AL 《The journal of physical chemistry. B》2006,110(37):18093-18096
Ultraviolet excitation of high surface area MgO films using 4.7 eV femtosecond pulses results in neutral Mg-atom desorption with hyperthermal kinetic energies in the range 0.1-0.4 eV. The Mg-atom hyperthermal energies and power dependences are similar to those previously observed using nanosecond pulsed excitation. Femtosecond two-pulse correlation measurements reveal the existence of different dynamical paths for Mg-atom desorption. One mechanism displays a sub-150 fs time scale and involves the simultaneous or near-simultaneous transition of two electrons to a 3-coordinated Mg(2+) site. Other paths display picosecond time scales that we associate with dynamics following electron trapping at 3-coordinated Mg(2+) surface sites. 相似文献
19.
Paquin R Ferrage F Mulder FA Akke M Bodenhausen G 《Journal of the American Chemical Society》2008,130(47):15805-15807
Side-chain carboxyl and carbonyl groups play a major role in protein interactions and enzyme catalysis. A series of (13)C relaxation experiments is introduced to study the dynamics of carboxyl and carbonyl groups in protein side chains on both fast (sub-ns) and slower (micros-ms) time scales. This approach is illustrated on the protein calbindin D(9k). Fast dynamics features correlate with hydrogen- and ion-binding patterns. We also identify chemical dynamics on micros time scales in solvent-exposed carboxyl groups, most probably due to exchange between the carboxylate and carboxylic acid forms. 相似文献