Water proton relaxation in solutions of serum albumin from healthy donors and patients with liver and renal failures

These studies demonstrate possible connection between a decreased affinity of human serum albumin (HSA) in patients with liver and renal failures and changes of the HSA hydration state. The relaxation times,T ₁ andT ₂, of water protons in aqueous protein solution depend on the interaction of water molecules with biopolymer macromolecules. We compared these relaxation times for aqueous solutions of HSA from healthy and sick donors. For latter the amount and correlation time of the bound water are higher than those in healthy donors HSA solutions. The influence of long-chain fatty acids on the albumin hydration was found to be small.     

Mobility and structure of human serum albumin powders in water-acetonitrile mixtures by1H NMR relaxation and FTIR spectroscopy

The effect of acetonitrile on protein dynamics was investigated for solid human serum albumin samples at various hydration levels. Temperature dependences of¹H nonselective nuclear magnetic resonanceT ₁ andT ₂ relaxation times at 27 MHz have been measured and data were interpreted in terms of three kinds of internal motions in the protein. Microdynamic parameters of the motions were obtained within a “model-free” approach. It was found that acetonitrile hardly affects the fast motions but noticeably influences the slow motion of side chain groups, shortening the correlation time and increasing the amplitude of the motion. The acetonitrile effect on dynamics is likely based on the appearance of additional free volume as a result of the formation of rigid helical parts in the protein structure. Water, plasticizing the protein structure, promotes the action of organic solvent. A definite part of side chain groups, slowly moving in the same frequency window as the rest of protein side chain groups, performs less constrained “liquidlike” motion. The relative population of these highly movable protons is closely correlated with the increment of the helical structure induced by water and acetonitrile.     

Response of lysozyme internal dynamics to hydration probed by13C and1H solid-state NMR relaxation

We have studied the hydration dependence of the internal protein dynamics of hen egg white lysozyme by naturally abundant¹³C and¹H nuclear magnetic resonance (NMR) relaxation. NMR relaxation timesT ₁, off-resonanceT _1p and proton-decoupled on-resonanceT _1p (only for carbon expriments) were measured in the temperature range from 0 to 50°C. The spectral resolution in carbon cross-polarization magic angle spinning spectrum allows to treat methine, methylene and methyl carbons separately, while proton experiments provide only one integral signal from all protons at a time. The relaxation times were quantitatively analyzed by the well-established correlation function formalism and model-free approach. The whole set of the data could be adequately described by a model assuming three types of motion having correlation times around 10^?4, 10^?9 and 10^?12 s. The slowest process originated from correlated conformational transitions between different energy minima, the intermediate process could be identified as librations within one energy minimum, and the fastest one is a fast rotation of methyl protons the symmetry axis of methyl groups. A comparison of the dynamic behavior of lysozyme and polylysine obtained from a previous study (A. Krushelnitsky, D. Faizullin, D. Reichert, Biopolymers 73, 1–15, 2004) reveals that in the dry state both biopolymers are rigid on both fast and slow time scales. Upon hydration, lysozyme and polylysine reveal a considerable enhancement of the internal mobility, however, in different ways. The side chains of polylysine are more mobile than those of lysozyme, whereas for the backbone a reversed picture is observed. This difference correlates with structural features of lysozyme and polylysine discussed in detail. Due to the presence of a fast spin diffusion, the analysis of proton relaxation data is a more difficult task. However, our data demonstrate that the correlation functions of motion obtained from carbon and proton experiments are substantially different. We explained this by the fact that these two types of NMR relaxation experiments probe the motion of different internuclear vectors. The comparison of the proton data with our previous results on proton relaxation timesT ₁ measured over a wide temperature range indicates that at low temperatures lysozyme undergoes structural rearrangements affecting the amplitudes and/or activation energies of motions.     

RSMR study of the hydration effect on the dynamics of some globular proteins

The dependence of the Mössbauer elastic scattering fraction on the hydration degree (h) has been studied for hydrated samples of human albumen (HSA), lysozyme and trypsin pancreatic inhibitor, within the range ofh between 0–0.75 g H₂O/g protein at 295 K, and for HSA with different hydration degrees (0.03, 0.25, 0.41, 0.65) in the temperature range between 100–320 K. An increase of the hydration degree forh>0.1 atT>200 K has been shown to result in the release of intramolecular mobility in proteins.     

Effect of the frequency of intramolecular motions on the NMR relaxation in liquid state temperature regime

Equations for the spectral densities of complex motion of a spin pair undergoing internal motion and isotropic/anisotropic overall rotation have been considered. The fluctuations of the interproton distances, caused by internal motion, have been taken into account in the theoretical equations. A method allowing a distinction between the isotropic and the anisotropic overall rotation of molecules has been proposed. The effect of the activation parameters of internal motions (known from the solid state study) on the measured T ₁ relaxation of the ¹³C and ¹H–¹H cross-relaxation rates has been analysed for methyl-β-D-galactopyranoside in DMSO-d6 solution. The conformational trans-gauche jumps of the methylene group are not fast enough to affect the T ₁ value of carbon C6 in the liquid state temperatures regime. Only the methyl group rotation is a very fast internal motion. This motion influences the carbon C7 relaxation and methyl protons–anomeric proton cross-relaxation. The values of interatomic distances between anomeric H(C1) and H(C5) as well as the three methyl protons H(C7) have been calculated from the cross-relaxation rates. The distance H(C1)–H(C7) fluctuates due to the rotation of methyl group. The application of the ‘model-free approach’ to study molecular dynamics in solutions is discussed.     

Rotational Inhibition and Magnetization Transfer in α-Crystallin Solutions

The magnetic field dependence (dispersion profile) of 1/T₁ of solvent water protons of solutions of the mammalian eye lens protein α-crystallin has a reversible nonlinear dependence on concentration which, as protein concentration increases above ∼20 wt.%, changes rapidly from a profile characteristic of mobile protein solute to a profile characteristic of rotationally immobilized protein (e.g., chemically cross-linked). From quantitative comparisons of new measurements of K at 200.1 MHz, the rate of water-to-protein interfacial magnetization transfer, with earlier data for α-crystallin and bovine serum albumin (BSA), we conclude, for α-crystallin, that: (i) Brownian rotation is slowed by intermolecular interactions at unexpectedly low concentrations; (ii) K is mediated by the newly reported, long-lived, protein hydration sites with the same surface density as BSA; and (iii) ¹⁴N peaks seen in solvent 1/T₁ arise from interfacial magnetization transfer plus diffusion to protein NH magnetization sinks.     

Exchange and dipolar spin-spin interaction and rotational diffusion in saturation transfer EPR spectroscopy

Saturation transfer EPR spectroscopy (STEPR) provides a means for investigating weak spin-spin interaction between spin-labelled molecules because the spectral intensity is proportional to the effective spin-lattice relaxation time,T ₁ ^eff. Rate equations for the spin population defferences yield equivalent results for the dependence ofT ₁ ^eff on the physical (or chemical) and Heisenberg spin exchange rates and show thatT ₁ ^eff depends on the extent of redistribution of saturation throughout the anisotropic spin label powder lineshape. This approach yields a particularly simple formulation for the dependence of the STEPR lineshape on slow rotational diffusion. The effects of spin exchange are readily distinguished from those of slow rotational diffusion because of the insensitivity of the STEPR lineshape in the former case. The characteristic dependence of the STEPR spectral intensity on spin concentration allows determination of the exchange rate and can be used for studying slow translational diffusion, e.g. of spin-labelled proteins. Dipolar relaxation induced by paramagnetic ions gives a linear dependence of the reciprocal spin label STEPR intensity on metal ion concentration. STEPR measurements with spin-labelled lipid molecules in gel phase membranes in the presence of Ni²⁺ ions yield reliable distance information and provide calibrations for use with other systems.     

A method for evaluating correlation times for tumbling and internal motion in macromolecules using cross-relaxation rate constants from proton NMR spectra

A method of estimating the correlation times and extent of internal motion of macromolecules using ¹H NMR is proposed. The method relies on measuring the cross-relaxation rate constant between resolved, identified protons separated by a fixed distance, for example 2, 3 protons of tyrosine residues or 4, 5 protons of tryptophan residues in proteins, and the 5, 6 protons of cytosine residues in DNA. For a rigid body, the cross-relaxation rate constant yields directly an estimate of the tumbling time. Deviation of its dependence on viscosity and temperature from expectations for a rigid body allows one to estimate the degree to which internal motions contribute to the relaxation. The method is illustrated for Ribonuclease A and a 20 base pair fragment of DNA corresponding to the trp operator of Escherichia coli. The calculated correlation time of RNAse A is about 8 ns at 298 K, in good agreement with expectations from hydrodynamic measurements. Tyrosine 25 has significant internal motion, characterized by an apparent amplitude of 50–60°, a correlation time of about 5 ns, and low activation energy. The correlation time of the fragment of DNA is about 6.4 ns at 298 K, in agreement with expectations for a rigid rod. The apparent activation energy was 3.8 kcal/mol, close to the value for the dependence of the viscosity of D₂O on temperature. Further, the same result was obtained regardless of the position of the base in the sequence, indicating that bending motions are of small amplitude on the nanosecond time scale for short fragments of DNA.     

Analysis of Polypeptide Backbone T1 Relaxation Data Using an Experimentally Derived Model

The time scale of internal motion of the Ala₃-Leu₄ peptide linkage plane of gramicidin A in its channel conformation is analyzed by interpreting solid-state ¹⁵N T₁ measurements taken at 4.7 and 9.4 T. The T₁ relaxation measurements have been interpreted using a structural model of local motion which has previously been experimentally determined. According to this model, the individual peptide plane motion is a rotational diffusion subject to a harmonic restoring potential about an axis through successive α carbons. The results which best fit the model to the data reported here and to previously determined information about the motions of the gramicidin channel suggest that the correlation time for librations of the Ala₃-Leu₄ peptide linkage is 36 ns. This suggests that the peptide backbone motions of the gramicidin channel may be on the same time scale as the translational motion of a cation in the channel.     

Nuclear quadrupole resonance and thermally activated molecular mobility in solids: Reorientations of symmetric molecular species and pseudorotation in trigonal-bipyramidal molecules

The specific features revealed in the behavior of temperature dependences of the spin-lattice relaxation time t ₁ of quadrupole nuclei involved in different types of thermally activated motions in crystals have been analyzed under the assumption that there exists a temperature dependence of the activation energy of motion. An expression for the spin-lattice relaxation rate has been obtained. This expression includes the effect under consideration and provides an adequate explanation for numerous anomalies in the behavior of the experimental dependences t ₁(T), which were revealed in the case of the motion of molecular fragments with a relatively large volume. A nontraditional method for evaluating the activation energy of motion under these conditions has been proposed.     

NMR spectral densities and two dimensional diffusion in TiS2(NH3)1.0 and TaS2(NH3)x

NMR measurements of proton spin-lattice relaxation times T₁ and T_1? in the layered intercalation compounds TiS₂(NH₃)_1.0 and TaS₂(NH₃)_x (x = 0.8, 0.9, 1.0) are reported as functions of frequency and temperature (100 K – 300 K). These observations probe the spectral density of magnetic fluctuations due to motions of the intercalated molecules at frequencies accessible to the T₁ (4–90 MHz) and T_1? (1–100 kHz) measurements. Since the average molecular hopping time (τ) can be changed by varying temperature, different regions of the spectral density can be examined. For T > 200 K, both T^?1₁ and T^?1_1? vary logarithmically with frequency, reflecting the two dimensional character of the molecular diffusion. The temperature dependence of T₁ suggests that a more accurate picture of the short time dynamics is required. No dependence of relaxation rate on vacancy concentration is found.     

Spin-lattice relaxation in multiple-pulse experiments as contrast parameter in NMR imaging of solids

The molecular motion contrast parameter for NMR imaging of solids and quasi-solids based on the spin-lattice relaxation (T _leff) in multiple-pulse experiments is discussed. For Ostroff-Waugh multiple-pulse sequence theT _leff contrast parameter is evaluated in slow and fast molecular motion regime and compared with spin-lattice relaxation in the rotating frame contrast parameter. It is shown thatT _leff is offering a good molecular motion contrast in NMR imaging of polymer systems. The radio-frequency pulse scheme forT _leff-imaging using magic-echo phase-encoding procedure for recording spatial distribution in solids is introduced. A method forT _leff-weighted imaging using gradient spin-echo valid for weak dipolar solids is also discussed. The one-dimensional protonT _leff image using Ostroff-Waugh pulse sequence in combination with frequency-encoding imaging procedure is presented for a phantom of poly(ethyleneoxide) and poly(methylmethacrylate). The distribution of mechanical stresses in a acrylate film on glass is investigated by protonT _leff-imaging. A proton spin-density image weighted byT _leff, for a mixture of two elastomers with different crosslink density is also shown.     

Model-Free Approach beyond the Borders of Its Applicability

Model calculations presented in this article show that commonly used methodology of¹⁵N relaxation data analysis completely fails in detecting nanosecond time scale motions if the major part of the molecule is involved in these motions. New criteria are introduced for the detection of such cases, based on the dependence of the apparent overall correlation time, derived from theT₁/T₂ratio, on the spectrometer frequency. Correctly estimating the overall rotation correlation time τ_Rwas shown to play the key role in model-free data analysis. It is found, however, that in cases of slow internal motions with characteristic times of more than 3–4 ns, the effective τ_Rprovided by theT₁/T₂ratio for individual amide nitrogens can be used for the characterization of the fast picosecond internal dynamics.     

Systematic bias in the model-free analysis of heteronuclear relaxation

Several theoretical and practical aspects of the use of heteronuclear relaxation to characterize the internal motion of biomacromolecules are examined. The treatment is cast in the terms of the popular model-free theory of G. Lipari and A. Szabo (J. Am. Chem. Soc.104, 4546 (1982)). In particular, the measurement of longitudinal relaxation in fully coupled systems is found to systematically bias obtained generalized order parameters. It is concluded that it is generally appropriate to utilize an experimental approach which ensures that heteronuclei are decoupled frorn bonded protons during sampling of relaxation. This restricts the types of experiments that can be used to sample relaxation by indirectly detected two-dimensional spectroscopy. The influence of a limited sampling of relaxation, typically required by practical limitations when two-dimensional spectroscopy is employed, is also examined. Finally, the relative importance of T₁ and NOE in determining model-free parameters is explored in conjunction with the impact of commonly used assumptions on the analysis.     

17O quadrupole resonance study of the ferroelectric phase transition in KH2PO4

The temperature dependence of the ¹⁷O NQR spectra in KH₂PO₄ has been measured using a proton-¹⁷O double resonance technique in the laboratory frame. The spectra are consistent with a model where the protons rapidly move above T_c between the two equilibrium sites in the O-H--O bonds, whereas the motion freezes in below T_c.     

7Li NMR study of the mobility of lithium ions in one-dimensional channels of Nb3Se4

The temperature dependence of the spin-lattice relaxation time T ₁ and the ⁷Li NMR spectra of the Li_0.7Nb₃Se₄ intercalation compound with one-dimensional channel structure have been studied. It is found that the temperature dependence of T ₁ exhibits two relaxation minima, and the quadrupole splitting in the Li NMR spectra shows an anomalous temperature behavior. The inference is drawn that the observed effects are associated with the high-rate diffusive motion of lithium ions along one-dimensional channels and the interchannel transitions.     

Molecular reorientations of 1-bromo- and 1-iodo-adamantanes 1H N.M.R. relaxation study

Second moments and spin-lattice relaxation times, T ₁ and T _1ρ, have been measured from 100 K to 400 K for the protons in powdered 1-bromo and 1-iodo-adamantanes. Analysis of these data have shown that the reorientations are uniaxial in the low temperature phases. In the high temperature disordered phase of bromo-adamantane, the reorientation is endospherical and a slow molecular translational motion also exists. In the high temperature disordered phase of iodo-adamantane the reorientation is 12-fold uniaxial, in agreement with the Incoherent Quasi-elastic Neutron Scattering (I.Q.N.S.) experiments. All the results correspond to the crystallographic structures deduced from X-ray scattering.     

Particle size dependence of relaxivity for silica-coated iron oxide nanoparticles

We investigate the particle size dependence of the relaxivity of hydrogen protons in an aqueous solution of iron oxide (Fe₃O₄) nanoparticles coated in silica for biocompatibility. The T₁ and T₂ relaxation times for various concentrations of silica-coated nanoparticles were determined by a magnetic resonance scanner. We find that the relaxivity increased linearly with increasing particle size. The T₂ relaxivity (R₂) is more than 50 times larger than the T₁ relaxivity (R₁) for the nanoparticle contrast agent, which reflects the fact that the T₂ relaxation is mainly influenced by outer sphere processes. The high R₂/R₁ ratio demonstrates that silica-coated iron oxide nanoparticles may serve as a T₂ contrast agents in magnetic resonance imaging with high efficacy.     

Proton spin-lattice relaxation study of the phase transition in TlH2AsO4

Measurements of the temperature and frequency dependence of the proton laboratory and dipolar frame spin-lattice relaxation rates in powdered TlH₂AsO₄ demonstrate that the ⁷⁵As quadrupole resonance frequency changes from v_Q ≈ 38 MHz at T ?T_c to v_Q 〈 10 MHz at T 〉 T_c. The structural phase transition at T_c = 251 K is thus connected with a disordering of the protons in the O–H … O bonds surrounding the AsO₄ groups.