Molecular Dynamics of Cyclodextrins in Water Solutions from NMR Deuterium Relaxation: Implications for Cyclodextrin Aggregation.

The aggregation of the most common natural cyclodextrins (α-, ß-, and γ-) in aqueous solutions is addressed by studying the CD-CD interactions using deuterium relaxation rates for deuterium labeled CDs. Relaxation times (T1) and their corresponding relaxation rates (R1 = 1/T1) provide information about the rotational correlation times of CDs and serve as a proxy for solute-solute interactions. Measured T1's for α-, ß-, and γ-CD at the lowest CD concentrations were in agreement with predictions of a hydrodynamic model for toroids, in particular with regard to the dependence of T1 on CD size. On the other hand, the dependence of T1's with respect to the increase in CD concentration could not be explained by hydrodynamic or direct interaction between CD molecules, and it is suggested that there is an equilibrium between monomeric and dimeric CD to account for the observed concentration dependence. No evidence in favor of large aggregates of CDs involving a non-negligible fraction was found for the investigated CDs.

The effect of the head-group spacer length of 12-s-12 gemini surfactants in the host-guest association with ß-cyclodextrin.

NMR spectroscopy has been used to study and characterize the interactions in solution between ß-CD and alkyl-α,ω-bis(dodecyldimethyl ammonium bromide) gemini surfactants with the following head-group spacer lengths: 2, 4, 6, 8, and 10. The application of the method of continuous variation gives as a result that 1:1 and 2:1 (ß-cyclodextrin-gemini) complexes are formed; the association stoichiometry is dependent on the spacer chain length, varying from 1.5 (for s=2) to 1.8 (for s=10). Assuming a two-step mechanism, the binding constants have been computed. In general, the overall binding constant slightly increases with an increase of the number of methylene groups in the spacer. The (1)H NMR spectra of the N-(CH(3))(2) groups in ß-cyclodextrin/gemini mixed solutions are split into two peaks for 12-10-12, suggesting that the gemini spacer can thread the ß-cyclodextrin so that the latter is positioned between the gemini head-groups. Inspection of the ROESY spectra allowed the establishment of several spatial proximities between the protons from the ß-CD and the gemini and for a spacer length of 10, the data indeed indicate that complexes are formed with the CD molecule positioned between the two charged head groups with the spacer passing through the CD molecule.

PFG-NMR diffusometry: a tool for investigating the structure and dynamics of noncommercial purified pig gastric mucin in a wide range of concentrations.

For the first time, Pulsed Field Gradient-Nuclear Magnetic Resonance, a powerful noninvasive tool for studying the dynamics and structure of complex gels, has been used to measure diffusion of probe molecules in aqueous solutions/gels of noncommercial purified pig gastric mucin (PGM), in a concentration range up to 5 wt %. Complementary data were obtained from rheology measurements. The combination of techniques revealed a strong pH dependency of the structure of the PGM samples while changes in concentration, ionic strength, and temperature appeared to induce less pronounced alterations. Viscosity was found to vary in a nonmonotonous way with pH, with the more viscous solutions found at intermediate pH. We propose that this finding is due to a reduced charge density at lower pH, which is expected to continuously increase the relative importance of hydrophobic associations. The results suggest a loose network of expanded fully charged PGM molecules with considerable mobility at neutral pH (pH 7.4). At intermediate pH (pH 4), a three-dimensional expanded network is favored. At pH 1, the charge density is low and microphase separation occurs since hydrophobic associations prevail. This leads to the formation of clusters concentrated in PGM molecules separated by regions depleted in PGM. The results obtained increase our knowledge about the gastric mucosal layer, which in vivo contains mucin in the same concentration range as that of the samples investigated here.

Macroscopic background gradient and radiation damping effects on high-field PGSE NMR diffusion measurements.

The effects of macroscopic background gradients due to susceptibility differences at the sample interfaces and of radiation damping on pulsed-gradient spin-echo (PGSE) experiments are examined. Both phenomena can lead to the seemingly strange effect of the echo signal growing as the gradient strength increases at low applied gradient strengths. For a freely diffusing species, background gradients manifest themselves as slight concave or convex inflections in the linearized PGSE attenuation curve, depending on the polarity of the applied gradient. The various means of overcoming macroscopic background gradient problems, including bipolar gradients, and their efficacy are examined experimentally and discussed. The effects of radiation damping can also result in the attenuation curve being nonlinear but, different from the effect of background gradients, the nonlinearity does not change with the polarity of the applied gradient. The vulnerability of the stimulated echo-based PGSE sequence and variations of Hahn-based PGSE sequences is investigated. Both background gradients and radiation damping have serious implications for accurate diffusion measurement determination.

Diffraction-like effects in a highly concentrated W/O emulsion: a PFG NMR study.

Diffraction-like effects have been observed by applying pulsed field gradient (PFG) nuclear magnetic resonance (NMR) to a highly concentrated water-in-oil (W/O) emulsion, made up of the nonionic surfactant C12E4 [CH3(CH2)11(OCH2CH2)4OH], n-decane, and brine [1 wt% NaCl(aq) solution]. The pulsed field gradient NMR data show one pronounced maximum and the shoulder of a second maximum in the attenuation curve of the NMR signal from water, the so-called Bragg interference peaks. From the diffraction-like peaks, the average distance (center to center) between the emulsion droplets can be obtained, in this case related to the average size of an emulsion droplet. Furthermore, we note that the long-term stability of the emulsion can be followed by pulsed field gradient NMR.

Exchange dynamics of surfactants in adsorption layers investigated by PFG NMR diffusion.

Surfactant adsorption layers on polystyrene latex particles are investigated concerning their equilibrium exchange dynamics with surfactant molecules in solution. Pulsed field gradient (PFG) nuclear magnetic resonance (NMR) self-diffusion measurements of the nonionic surfactant C12(EO)5 are performed, making use of the difference of the diffusion coefficients of adsorbed and free surfactant in solution. On variation of the gradient pulse spacing the echo decay data show a transition from fast to slow exchange, giving the possibility of a quantitative evaluation. A two-site model is applied to fit the data and obtain average residence times of the surfactant in both sites. The results are consistent with diffusion-controlled adsorption of surfactant monomers and additional micellar contributions to the exchange, whereas no evidence for kinetic barriers is found for these nonionic surfactants on the relevant timescale (10 ms).

An NMR Self-Diffusion Investigation of Aggregation Phenomena in Solutions of Ethyl(hydroxyethyl)cellulose.

Water solutions of ethyl(hydroxyethyl)cellulose have been investigated with the PFG NMR technique and by means of 1H NMR relaxation measurements. The echo decays in PFG experiments have been recorded for EHEC with concentrations ranging from the dilute (c < c) to the semidilute (c > c). In addition, the temperature dependence of the echo decay was investigated. It is shown that the echo decays deviate substantially from Gaussian diffusion behavior. We discuss this in terms of polydispersity, aggregation, and scaling effects. It is argued that the observed deviations are too large to be caused by polydispersity effects alone. We base this conclusion on the observation that the molar mass scaling of the polymer diffusion coefficient goes roughly as M-4. We focus on using a log-normal distribution of diffusion coefficients in the evaluation procedure of the PFG data. It is found that by comparing the diffusion coefficients extracted from this type of procedure with "Dmean" as extracted from a "stretched exponential" approach, there is a difference in the evaluated diffusion constant by up to 1 order of magnitude. The difference depends on the degree of deviation from linear behavior in the echo decay when represented in a logarithmic plot vs the relevant experimental parameter.

Dynamic properties of the backbone of an integral membrane polypeptide measured by 2H-NMR.

The 2H-NMR spectrum of the exchangeable hydrogens of the synthetic amphiphilic polypeptide, lys2-gly-leu24-lys2-ala-amide, was measured for the solid peptide at room temperature and, as a function of temperature, for the peptide incorporated into hydrated dipalmitoylphosphatidylcholine (DPPC) bilayers. This study is a prototype of a similar class of experiments which can be carried out on integral membrane proteins to characterize, quantitatively, the dynamic properties of integral membrane proteins. At temperatures below the DPPC gel-liquid crystalline phase transition, the 2H NMR spectrum was very similar to that of the solid peptide indicating that the peptide was immobilized in the lipid bilayer on the time scale (approximately equal to 10(-5) s) of the 2H-NMR measurements. The 2H-NMR spectrum above the phase transition corresponded to that expected from a peptide in the alpha-helical conformation reorienting rapidly about the symmetry axis of the alpha-helix. Measurements of the quadrupolar echo relaxation time, T2e, gave a quantitative measure of the correlation time, tau c, for this motion. The value of tau c decreased rapidly with increasing temperature as the fraction of DPPC molecules in the liquid crystalline phase increased, reaching a value of 2 X 10(-7) s above the phase transition. The observation of a characteristic minimum in T2e as the temperature was raised provided a definitive, quantitative interpretation of the T2e measurements. Using the known geometry of the peptide and the theory of uniaxial rotational diffusion, a value of eta = 1.1 poise was obtained for the effective viscosity of the membrane in close agreement with values obtained previously from transient linear dichroism measurements.

The interactions between monovalent ions and phosphatidyl cholines in aqueous bilayers.

The interactions of lithium and sodium ions and water with phosphatidylcholine bilayers have been studied by means of 7Li, 23Na and 2H NMR quadrupole splittings. The experimental results are interpreted in terms of a simple three-site model with two anisotropic sites ('binding' sites) and one isotropic site ('free' ions and water molecules). The findings obtained for the zwitterionic model membrane are compared with previous investigations of lamellar phases composed of ionic and nonionic amphiphiles. It is shown that the data obtained are compatible with our previous suggestion [Lindblom, G., Persson, N.-O., and Arvidson G. (1976) Adv. Chem. Ser. 152, 121] that an increase in the salt content in the water layer induces a conformational change in the polar head group of phosphatidylcholine. Thus at high salt concentration the phosphocholine head group tends to be oriented perpendicular to the lipid bilayer surface. The study also shows that increasing the amount of salt leads to a squeezing out of water between the bilayers. This is interpreted in terms of a reduction of the repulsion forces between the bilayers.

Molecular organization in the liquid--crystalline phases of lecithin--sodium cholate-water systems studied by nuclear magnetic resonance.

The molecular organization in the hexagonal and lamellar phases of the ternary systems lecithin--sodium cholate--water has been investigated by using a variety of nuclear magnetic resonance techniques. The main findings and conclusions are the following: (i) When calculated on a mole fraction basis, the phase equilibria are insensitive to changes in the alkyl chains of the lecithin. (ii) When incorporated into a lecithin bilayer, cholate exerts a strong perturbation on the lecithin alkyl chain order, giving a large decrease of the order parameters. (iii) This decrease of the order occurs since the average cross-sectional area per alkyl chain increases probably as a result of cholate placing itself flat on the bilayer surface. (iv) The diffusion of lecithin molecules is approximately equally rapid in the lamellar and hexagonal phases. (v) The hexagonal phase is formed by rodlike aggregates with the polar groups at the surface of the rods and with a continuous hydrocarbon core. The rods are not formed by stacking disklike mixed micelles. (vi) With the interpretations of the molecular packing and the phase structures, the observed phase equilibria are in good agreement with current theories of the factors that govern phase behavior in amphiphile--water systems.