Proton magnetic relaxation process during the polymerization of hemoglobin S

The proton spin-lattice and spin-spin magnetic relaxation times are investigated at 4 MHz in samples of hemoglobin A and S with intracellular concentrations, at 36 °C, and during spontaneous deoxygenation. Magnetic relaxation behaves differently in the solutions of hemoglobin A and S. The possible causes of this behavior are discussed: changes in molecular mobility, the variations of hemoglobin magnetism, and the appearance of microinhomogeneities in the solutions of hemoglobin S at the end of the polymerization process.     

The line shapes of the water proton resonances of red blood cells containing carbonyl hemoglobin, deoxyhemoglobin, and methemoglobin: implications for the interpretation of proton MRI at fields of 1.5 T and below

The 300 MHz (7 T) water proton resonances of suspensions of red blood cells containing paramagnetic deoxyhemoglobin or methemoglobin can be resolved into two broad lines assignable to intra- and extracellular water which undergoes rapid T2 relaxation by diffusion in magnetic field gradients induced by the intracellular paramagnets. The width of the resolved lines allowed an estimate of the maximum contribution that diffusion makes to T2 relaxation at 7 T. The dependence of the diffusion contribution on the square of the strength of the static magnetic field suggest that diffusion makes a small contribution to water proton T2 relaxation at 1.5 T compared to 7 T, and a negligible one at 0.5 T in early and intermediate hematomas containing deoxyhemoglobin or methemoglobin in intact red blood cells. At the lower field strengths, water proton T2 relaxation is apparently dominated by the rapid chemical exchange (mean lifetime tau = 10 msec) between the intra- and extracellular environments.     

Consequences of (129)Xe-(1)H cross relaxation in aqueous solutions.

We have investigated the transfer of polarization from (129)Xe to solute protons in aqueous solutions to determine the feasibility of using hyperpolarized xenon to enhance (1)H sensitivity in aqueous systems at or near room temperatures. Several solutes, each of different molecular weight, were dissolved in deuterium oxide and although large xenon polarizations were created, no significant proton signal enhancement was detected in l-tyrosine, alpha-cyclodextrin, beta-cyclodextrin, apomyoglobin, or myoglobin. Solute-induced enhancement of the (129)Xe spin-lattice relaxation rate was observed and depended on the size and structure of the solute molecule. The significant increase of the apparent spin-lattice relaxation rate of the solution phase (129)Xe by alpha-cyclodextrin and apomyoglobin indicates efficient cross relaxation. The slow relaxation of xenon in beta-cyclodextrin and l-tyrosine indicates weak coupling and inefficient cross relaxation. Despite the apparent cross-relaxation effects, all attempts to detect the proton enhancement directly were unsuccessful. Spin-lattice relaxation rates were also measured for Boltzmann (129)Xe in myoglobin. The cross-relaxation rates were determined from changes in (129)Xe relaxation rates in the alpha-cyclodextrin and myoglobin solutions. These cross-relaxation rates were then used to model (1)H signal gains for a range of (129)Xe to (1)H spin population ratios. These models suggest that in spite of very large (129)Xe polarizations, the (1)H gains will be less than 10% and often substantially smaller. In particular, dramatic (1)H signal enhancements in lung tissue signals are unlikely.     

H,N,C-triple resonance NMR of very large systems at 900 MHz

We provide quantitative signal to noise data and feasibility study at 900 MHz for 1H-15N-13C triple resonance backbone assignment pulse sequences obtained from a medium sized 2H, 13C, 15N labeled protein slowed down in glycerol-water solution to mimic relaxation and spectroscopic properties of a much larger protein system with macromolecular tumbling correlation time of 52 and 80 ns, respectively, at 296 and 283 K (corresponding to molecular weights of 130 and 250 kDa). Comparisons of several different schemes for transferring magnetization from proton to nitrogen and back to proton confirms Yang and Kay's 1999 prediction that avoiding the unfavorable relaxation properties of 1H-15N multiple quantum coherence in the TROSY phase cycle of the final 15N-1H transfer before acquisition is crucial for maximal sensitivity from these very large molecular weight systems. We also show results which confirm some predictions regarding the superiority of TROSY at 900 MHz vs. 800 MHz especially as the molecular weights become very large.     

1H,15N,13C-triple resonance NMR of very large systems at 900 MHz

We provide quantitative signal to noise data and feasibility study at 900 MHz for 1H-15N-13C triple resonance backbone assignment pulse sequences obtained from a medium sized 2H, 13C, 15N labeled protein slowed down in glycerol-water solution to mimic relaxation and spectroscopic properties of a much larger protein system with macromolecular tumbling correlation time of 52 and 80 ns, respectively, at 296 and 283 K (corresponding to molecular weights of 130 and 250 kDa). Comparisons of several different schemes for transferring magnetization from proton to nitrogen and back to proton confirms Yang and Kay's 1999 prediction that avoiding the unfavorable relaxation properties of 1H-15N multiple quantum coherence in the TROSY phase cycle of the final 15N-1H transfer before acquisition is crucial for maximal sensitivity from these very large molecular weight systems. We also show results which confirm some predictions regarding the superiority of TROSY at 900 MHz vs. 800 MHz especially as the molecular weights become very large.     

Extensive NMRD studies of Ni(II) salt solutions in water and water-glycerol mixtures

Aqueous solutions of simple nickel(II) salts are a classical test case for theories of the paramagnetic relaxation enhancement (PRE) and its dependence on the magnetic field (nuclear magnetic relaxation dispersion, NMRD), going back to late fifties. We present here new experimental data, extending the NMRD range up to 21T (900 MHz). In addition to salt solutions in (acidified) water, we have also measured on solutions containing glycerol. The aqueous solution data do not show any significant changes compared to the earlier experiments. The interpretation, based on the general ("slow-motion") theory is also similar to the earlier work from our laboratory. The NMRD-data in mixed solvents are qualitatively different, indicating that the glycerol not only changes the solution viscosity, but may also enter the first coordination sphere of the metal ion, resulting in lower symmetry complexes, characterized by non-vanishing averaged zero-field splitting. This hypothesis is corroborated by molecular dynamics simulations. A strategy appropriate for interpreting the NMRD-data for the chemically complicated systems of this type is proposed.     

In vivo MR evaluation of Gd-DTPA conjugated to dextran in normal rabbits.

A dextran-Gd-DTPA compound has been recently synthesized utilizing 70,800 Da molecular weight dextran. This polymeric contrast agent for magnetic resonance imaging has been found chemically to be very stable and to demonstrate in vitro improved relaxivities of 1.5 to 2.3 times that of monomeric Gd-DTPA at 100 MHz. This MR experiment examines the in vivo distribution and relaxivity of the 70,800 Da molecular weight dextran-Gd-DTPA compound in a rabbit model by measuring the change in signal-to-noise ratio of a variety of organs (renal cortex, renal medulla, liver, brain, and torcula herophile) compared to the preinjection state. Results demonstrate prolonged (beyond 60 min) enhancement of the renal cortex, renal medulla, liver and torcula, and no enhancement of brain parenchyma.     

Relaxation of protons by radicals in rotationally immobilized proteins

Proton spin-lattice relaxation by paramagnetic centers may be dramatically enhanced if the paramagnetic center is rotationally immobilized in the magnetic field. The details of the relaxation mechanism are different from those appropriate to solutions of paramagnetic relaxation agents. We report here large enhancements in the proton spin-lattice relaxation rate constants associated with organic radicals when the radical system is rigidly connected with a rotationally immobilized macromolecular matrix such as a dry protein or a cross-linked protein gel. The paramagnetic contribution to the protein-proton population is direct and distributed internally among the protein protons by efficient spin diffusion. In the case of a cross-linked-protein gel, the paramagnetic effects are carried to the water spins indirectly by chemical exchange mechanisms involving water molecule exchange with rare long-lived water molecule binding sites on the immobilized protein and proton exchange. The dramatic increase in the efficiency of spin relaxation by organic radicals compared with metal systems at low magnetic field strengths results because the electron relaxation time of the radical is orders of magnitude larger than that for metal systems. This gain in relaxation efficiency provides completely new opportunities for the design of spin-lattice relaxation based contrast agents in magnetic imaging and also provides new ways to examine intramolecular protein dynamics.     

Concentration Dependence of NMR T1 in Agar Solutions

In this study, in order to explain solvent proton relaxation mechanism, the spin-lattice relaxation time (T1) of agar solutions was measured as a function of agar concentration. Relaxation measurements were carried out by a FT-NMR spectrometer operating at 60 MHz and inversion recovery pulse squence was used. Relaxation rate(1/T1a) was linearly proportional to concentration of agar solution (C), and the T1 mechanism of solvent water protons in agar solutions should be caused by the chemical exchange of water protons between free and bound water.     

Concentration Dependence of NMR T1 in Agar Solutions

In this study, in order to explain solvent proton relaxation mechanism, the spin-lattice relaxation time (T1) of agar solutions was measured as a function of agar concentration. Relaxation measurements were carried out by a FT-NMR spectrometer operating at 60 MHz and inversion recovery pulse squence was used. Relaxation rate(1/T1a) was linearly proportional to concentration of agar solution (C), and the T1 mechanism of solvent water protons in agar solutions should be caused by the chemical exchange of water protons between free and bound water.     

General features of proton longitudinal relaxation in proteins in solution

Time courses of the recovery upon nonselective inversion of all individual proton magnetizations in several globular proteins in aqueous (²H₂O) solution were calculated for varying degrees of rotational correlation time of the molecule (10⁻⁹ s ∼ ∞) and compared with the experimental data on various proteins at 400 MHz. In the calculation, the spinrelaxation mechanism was assumed to be solely the dipolar interaction between protons, and the three-site random jumps of the methyl groups, along with the rotation of the whole molecule, were taken into account. The following conclusions were drawn. ( 1 ) For proteins whose molecular weights are below ∼ 10,000, whole-molecule rotation is a dominant source of relaxation, and the longitudinal relaxation times may vary considerably from proton to proton. (2) For proteins whose molecular weights are above ∼20,000, methyl group rotations assisted by spin diffusion are common and major sources of relaxation, producing T₁ values close to 1 s. In the intermediate region (molecular weight 10,000 ∼ 20,000), both whole-molecule rotation and methyl group rotations contribute significantly to relaxation. (3) In some proteins, segmental motions are as important as methyl group rotations in determining relaxation rate.     

Stereospecific Assignment of the Asparagine and Glutamine Side Chain Amide Protons in Random-Coil Peptides by Combination of Molecular Dynamic Simulations with Relaxation Matrix Calculations

A database with correct stereospecific assignment is also important for accurate chemical shift prediction. Here, usually chemical shifts of random-coil model peptides or from a protein database are used for correct parameterization. However, an analysis of the data stored in the Biological Magnetic Resonance Data Bank (BMRB) shows that a substantial part of all stereospecific assignments stored is not correct. Here, we provide the stereospecific assignment of amide side chain protons in the amino acids glutamine and asparagine in the model peptides Gly-Gly-Asn-Ala-NH₂ and Gly-Gly-Gln-Ala-NH₂. Nuclear Overhauser enhancement spectroscopy (NOESY) spectra were back-calculated with the full relaxation matrix formalism implemented in AUREMOL-RELAX from molecular dynamics trajectories created with GROMACS and compared with experimental nuclear magnetic resonance spectra measured at 800 MHz proton resonance frequency. The comparison of simulated with experimental NOESY spectra permitted the unambiguous stereospecific assignment of the side chain amide and H^β protons of asparagine and glutamine in the random-coil peptides.     

The influence of macromolecular polymerization of spin-lattice relaxation of aqueous solutions

The docking or polymerization of globular proteins is demonstrated to cause changes in proton NMR spin-lattice (T1) relaxation times. Studies on solutions of lysozyme, bovine serum albumin, actin, and tubulin are used to demonstrate that two mechanisms account for the observed changes in T1. Polymerization displaces the hydration water sheath surrounding globular proteins in solution that causes an increase in T1. Polymerization also slows the average tumbling rate of the proteins, which typically causes a contrary decrease in T1. The crystallization reaction of lysozyme in sodium chloride solution further demonstrates that the "effective" molecular weight can either decrease or increase T1 depending on how much the protein is slowed. The displacement of hydration water increases T1 because it speeds up the mean motional state of water in the solution. Macromolecular docking typically decreases T1 because it slows the mean motional state of the solute molecules. Cross-relaxation between the proteins and bound water provides the mechanism that allows macromolecular motion to influence the relaxation rate of the solvent. Fast chemical exchange between bound, structured, and bulk water accounts for monoexponential spin-lattice relaxation. Thus the spin-lattice relaxation rate of water in protein solutions is a complex reflection of the motional properties of all the molecules present containing proton magnetic dipoles. It is expected, as a result, that the characteristic relaxation times of tissues will reflect the influence of polymerization changes related to cellular activities.     

Ultrasonic spectroscopy in bovine serum albumin solutions

Ultrasonic absorption and velocity spectra in bovine serum albumin (BSA) aqueous solutions have been measured at 20 degrees C over the broad frequency range 0.1-1600 MHz in the pH range 1.5-13.2. Five different techniques were used: the plano-concave resonator, plano-plano resonator, pulse-echo overlap, Bragg reflection, and high-resolution Bragg reflection methods. The absorption spectrum at neutral pH was well fitted to the relaxation curve assuming a distribution of relaxation frequency with a high-frequency cutoff and long low-frequency tail. The relaxation behavior was interpreted in terms of various degrees of hydration of BSA molecules. At acid pH's, excess absorption over that at pH 7 was explained by double relaxation. The pH dependences of the relaxation frequency and maximum absorption per wavelength showed that the relaxation at about 200 kHz was related to the expansion of molecules and that at 2 MHz resulted from the proton transfer reaction of carboxyl group. At alkaline pH's, the excess absorption was explained by triple relaxation. The relaxation at about 200 kHz was associated with a helix-coil transition, and the two relaxations at 2 and 15 MHz were attributed to the proton transfer reactions of phenolic and amino groups, respectively. The rate constants and volume changes associated with these processes were estimated.     

The use of fast field cycling to evaluate the time domain relaxation of starches from tropical fruit seeds

ABSTRACT

A proton NMR relaxation study of the molecular dynamics in flour samples of ‘Eugenia uniflora’ (‘Pitanga’), ’Citrus reticulate’ (‘Tangerine’), ‘Prunus persica’ (‘Peach’), ‘Vitis vinifera’ (‘Grape’), and ‘Cucumis melo’ (‘Honeydew melon’) seeds is presented. The spin–lattice relaxation time was obtained over a broad frequency range from 100?kHz to 100?MHz using both conventional and fast field-cycling NMR techniques. This relaxometry study made possible a comparison between the molecular dynamics behaviour of starch from different fruit seeds and from potatoes. I was possible to conclude that the spin–lattice dispersion presents slightly differences for all samples, in particular at low frequencies. All relaxation dispersions could be well interpreted in terms of power-law relaxation models and domains with different power-law relaxation exponents. For the Peach seeds flour the relaxation dispersion at low frequencies was very similar to that observed for potato’s starch incorporated with 5% organoclay Viscogel B8 nanoparticles.     

Relaxation of solvent protons and deuterons by protein-bound Mn2+ ions. Theory and experiment for Mn2+-concanavalin A

Despite the demonstrated utility of measurements of the magnetic field dependence of the magnetic relaxation rates of solvent protons in solutions of metalloproteins as an indicator of biochemical changes, it is becoming increasingly evident that quantitative comparisons of such data with the theory of relaxation, limited by the approximations and assumptions usually made, yield results for the strength of the solvent-paramagnetic ion interaction that generally do not make chemical sense. These results, when expressed as the number of solvent-donated ligands of the ions, usually give too large a value, typically by about twofold. It has been suggested by several investigators that a comparison of proton and deuteron relaxation rates could resolve the problem. Data are presented for the longitudinal relaxation rates of solvent protons and deuterons over more than four decades of magnetic field (from 0.01 to 270 MHz proton Larmor frequency) for solutions of Mn²⁺-concanavalin A, a protein for which the physical biochemistry is thoroughly documented, one that should be particularly tractable for such comparisons. The main conclusion is that, in the general case, there is no decade of magnetic field over which the mathematical criterion of best agreement of data with theory can be relied upon to yield quantitatively correct biochemical results; rather, biochemistry must still be a guide for elucidating relaxation pathways.     

Proton magnetic relaxation in solutions of E. coli ribosomal RNA containing Mn2+ ions

A study has been made over a range of temperatures and magnetic field strengths of the spin relaxation of water protons in aqueous solutions of E. coli ribosomal RNA containing Mn²⁺ ions. The effects of the paramagnetic ions are enhanced in the presence of the RNA. As the temperature falls T ₁ passes through a minimum value, the magnitude of which is field dependent, and this is attributed to a change in dipolar relaxation mechanism from rotation of the aquocomplex to electron spin relaxation. The relevance of this work is assessed in relation to other work on proton relaxation enhancement in Mn²⁺-containing solutions of biopolymers.     

NMR relaxation dispersion of vulcanized natural rubber

The dependence of the 1H spin-lattice relaxation time on the magnetic field strength has been determined for linear and cross-linked polyisoprene for Larmor frequencies between 5 kHz and 20 MHz. Universal power-law relations are found for all temperatures and cross-link densities under investigation and are compared to published results of rotating-frame experiments on similar natural rubber samples. The shape of the individual dispersion functions can be superposed into a master curve using appropriate shift factors. While addition of filler particles even at large weight fractions has only a minor effect on the relaxation times, uniaxial deformation and swelling are demonstrated to alter the molecular dynamics significantly.     

Monitoring the Air Influence on Cement–Lime Mortar Hydration Using Low-Field Nuclear Magnetic Resonance Relaxometry

In the present study, we investigate the relationship between the nuclear magnetic resonance relaxation rate and the hydration time in two types of masonry cement-lime mortar. The studies are performed with the mortars both in an enclosed and a standard atmosphere to monitor the air influence on cement-lime mortar hydration and setting. The constituents of the investigated mortar samples are: cement, slaked lime, sand and water. They were mixed to achieve a flow spread of 10?cm. These types of mortars are usually suitable for historical masonry maintenance, but they can also be used for modern buildings, or even for concrete structures coatings to prevent concrete carbonation. All nuclear magnetic resonance relaxation experiments were performed at 20?°C using a low-field nuclear magnetic resonance instrument operable at 20?MHz proton resonance frequency. A slowing down of the hydration kinetics is demonstrated for the samples kept in closed atmosphere conditions. The results contribute to the understanding of cement–lime mortar hydration, carbonation and setting under closed atmosphere conditions.     

Dielectric response of water confined in metal–organic frameworks

Dielectric response of water confined in metal–organic frameworks was investigated in broad temperature range from 140 to 410 K and from 20 Hz to 1 MHz using a capacitance bridge. Several dispersion regions of characteristic shape were found, caused by freezing–melting of adsorbed water molecules, which disappear after a prolonged heating at 410 K. Temperature dependencies of relaxation time of confined water molecules were obtained and are compared to those of water confined in MCM-41 mesoporous molecular sieves.