Neutron Spin Echo Spectroscopy Research Articles

Li+Transport in Poly(Ethylene Oxide) Based Electrolytes: Neutron Scattering, Dielectric Spectroscopy, and Molecular Dynamics Simulations

The dynamics of Li(+) transport in polyethylene oxide (PEO) and lithium bis(trifluoromethanesulfonyl)imde mixtures are investigated by combining neutron spin-echo (NSE) and dielectric spectroscopy with molecular dynamics (MD) simulations. The results are summarized in a relaxation time map covering wide ranges of temperature and time. The temperature dependence of the dc conductivity and the dielectric α relaxation time is found to be identical, indicating a strong coupling between both. The relaxation times obtained from the NSE measurements at 0.05 Å(-1)<q<0.2 Å(-1) are of similar magnitude as the relaxation time of Li(+) predicted by MD simulation. Our results suggest that the characteristic live times of the ions within the oxygen cages are mainly determined by the α relaxation that corresponds to local segmental motions of polymers, to a much lesser extent by the main chain relaxation, and not at all by the β relaxation or other faster processes. It is the first time decisive experimental evidence for a microscopic picture of the Li ion transportation process is shown in which the PEO chain forms EO cages over several monomer units and the Li ion "jump" from cage to cage. The role of the backbone of the polymer is discussed and contributes signifcantly to the Li ion transportation process. Moreover, detailed characteristic length and time scales of the Li(+) transport process in this polymer electrolyte are identified and interpreted.

Nanoscale protein dynamics: A new frontier for neutron spin echo spectroscopy

Recent studies show that neutron spin echo spectroscopy (NSE) can reveal long-range protein domain motions on nanometer lengthscales and on nanosecond to microsecond timescales. This unique capability of NSE provides new opportunities to understand protein dynamics and functions, such as how binding signals are propagated in a protein to distal sites. Here we review our applications of NSE to the study of nanoscale protein domain motions in a set of cell signaling proteins. We summarize the theoretical framework we have developed, which allows one to interpret the NSE data (Biophys. J. 99, 3473 (2010) and Proc. Natl. Acad. Sci. USA 102, 17646 (2005)). Our theoretical framework uses simple concepts from nonequilibrium statistical mechanics, and does not require elaborate molecular dynamics simulations, complex fits to rotational motion, or elastic network models. It is thus more robust than multiparameter techniques that require untestable assumptions. We also demonstrate our experimental scheme involving deuterium labeling of a protein domain or a subunit in a protein complex. We show that our selective deuteration scheme can highlight and resolve specific domain dynamics from the abundant global translational and rotational motions in a protein. Our approach thus clears significant hurdles to the application of NSE for the study of protein dynamics in solution.

Influence of charge density on bilayer bending rigidity in lipid vesicles: A combined dynamic light scattering and neutron spin-echo study

We report a combined dynamic light scattering and neutron spin-echo study on vesicles composed of the uncharged stabilizing lipid 1,2-dimyristoyl-sn-glycero-3-phosphatidylcholine (DMPC) and the cationic lipid 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP). Mechanical properties of a model membrane and thus the corresponding bilayer undulation dynamics can be specifically tuned by changing its composition through lipid headgroup or acyl chain properties. We compare the undulation dynamics in lipid vesicles composed of DMPC/DOTAP to vesicles composed of a mixture of the uncharged helper lipid DMPC with the also uncharged reference lipid 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC). We have performed dynamic light scattering on the lipid mixtures to investigate changes in lipid vesicle size and the corresponding center-of-mass diffusion. We study lipid translational diffusion in the membrane plane and local bilayer undulations using neutron spin-echo spectroscopy, on two distinct time scales, namely around 25 ns and around 150 ns. Finally, we calculate the respective bilayer bending rigidities κ for both types of lipid vesicles. We find that on the local length scale inserting lipid headgroup charge into the membrane influences the bilayer undulation dynamics and bilayer bending rigidity κ less than inserting lipid acyl chain unsaturation: We observe a bilayer softening with increasing inhomogenity of the lipid mixture, which could be caused by a hydrophobic mismatch between the acyl chains of the respective lipid components, causing a lateral phase segregation (domain formation) in the membrane plane.

Direct Observation of Nonaffine Tube Deformation in Strained Polymer Networks

We present a one-to-one comparison of polymer segmental fluctuations as measured by small angle neutron scattering in a network under deformation with those obtained by neutron spin echo spectroscopy. This allows an independent proof of the strain dependence of the chain entanglement length. The experimentally observed nonaffine square-root dependence of the tube channel on strain is in excellent agreement with theoretical predictions and permits us to exclude an often invoked nondeformed as well as affinely deformed tube.

Nanoscale Protein Dynamics and Long-Range Allostery in Cell Signaling

An emerging point of view in protein chemistry is that proteins are not the static objects that are displayed in textbooks but are instead dynamic actors. Protein dynamics plays a fundamental role in many diseases, and spans a large hierarchy of timescales, from picoseconds to milliseconds or even longer. Nanoscale protein domain motion on length scales comparable to protein dimensions is key to understanding how signals are relayed through multiple protein-protein interactions. A canonical example is how the scaffolding proteins NHERF1 and ezrin work in coordination to assemble crucial membrane complexes. As membrane-cytoskeleton scaffolding proteins, these provide excellent prototypes for understanding how regulatory signals are relayed through protein-protein interactions between the membrane and the cytoskeleton. Here, we review recent progress in understanding the structure and dynamics of the interaction. We describe recent novel applications of neutron spin echo spectroscopy (NSE) to reveal the dynamic propagation of allosteric signals by nanoscale protein motion, and present a framework to analyze the NSE data.

Experimental determination of bending rigidity and saddle splay modulus in bicontinuous microemulsions

Elastic properties of surfactant membranes can be described in terms of the bending rigidity κ and the saddle splay modulus . Phase diagram measurements and neutron scattering experiments allowed the determination of these parameters. Recent simulations showed that the bending rigidity, which is deduced from the characteristic length scales in the microemulsion, is a mixture of and κ. By combining neutron spin echo (NSE) spectroscopy, small angle neutron scattering (SANS) and phase diagram measurements, we show that also experimentally the different contributions can be separated. For supercritical CO2 microemulsions and bicontinuous microemulsions with additives, the prefactors of the and κ contributions are determined and compared to those from simulations.

Steady State Dynamics of Enzyme Catalytic Action in Solution

Wide-angle X-ray scattering (WAXS) and neutron spin-echo (NSE) spectroscopy were used to study of the dynamics of adenylate kinase (Adk) as it undergoes steady state catalytic cycling. Analysis of the WAXS data indicates that it cannot be completely explained as a combination of open and closed states and suggests that ADP-bound Adk in solution mostly populates a closed conformation as demonstrated in earlier NMR studies [1,2,3]. The presence of other intermediates must be hypothesized to explain the observed data. Correlated internal dynamics on ps-ns timescales for the protein during active cycling appear to be slower than comparable dynamics in the open and closed states at certain length scales and faster at others. Our NSE results on active enzyme catalysis are consistent with the picture of energetic counterweight balancing on substrate binding. The enzyme appears to shift mobile regions during active catalysis for subsequent substrate release [4]. These methods will help define functional motions in proteins. Supported by DOE Great Lakes Bioenergy Research Center (DOE BER Office of Science DE-FC02-07ER64494)[1] Henzler-Wildman, K.A., V. Thai, M. Le1, M. Ott, M. Wolf-Watz, T. Fenn, E. Pozharski, M. A. Wilson, G. A. Petsko, M. Karplus, C. G. Hubner, and D. Kern (2007b) Intrinsic motions along an enzymatic reaction trajectory. Nature 450, 838-844.[2] Jorgen Aden and Magnus Wolf-Watz, (2007) NMR Identification of Transient Complexes Critical to Adenylate Kinase Catalysis J. AM. CHEM. SOC., 129, 14003-14012[3] M. D. Daily, L. Makowski, G. N. Phillips, and Q. Cui, (2012) Large-scale motions in the adenylate kinase solution ensemble: Coarse-grained simulations and comparison with solution X-ray scattering, Chem. Phys. 396, 84-91.[4] Muller, C.W., Schlauderer, G.J., Reinstein, J. & Schulz, G.E. (1996). Adenylate kinase motions during catalysis, an energetic counterweight balancing substrate binding. Structure 4, 147-156.

A microscopic view on the large scale chain dynamics in nanocomposites with attractive interactions

We use neutron spin-echo spectroscopy to investigate the large scale chain dynamics in unentangled polymer nanocomposites where stable polymer layers around nanoparticles are dynamically formed due to attractive segment–surface interactions. The work here focuses on the detailed microscopic characterization of the dynamics within these layers of bound poly(ethylene glycol) (PEO) and poly(butylene oxide) (PBO) chains at a fixed silica nanoparticle fraction of 15%. The substitution of hydroxy by methoxy terminated chains thereby clearly evidences the importance of the chain end chemistry in these systems as the layer structure and dynamics therein significantly depend on the specific interaction mechanism. The experimental data reveal a densely packed thick shell of end-attached chains in the case of hydroxy ends contrasted by a thin shell of laterally adsorbed chains with multiple attachments in the methoxy case. In all cases a consistent quantitative modeling is presented that evidences unchanged segmental dynamics within the bound layers. The obtained picture is further validated on an independent model system based on PBO polymers which shows surprisingly similar chain dynamics as for PEG in the nanocomposite pointing to a very generic dynamic scenario.

Relationship between mesoscale dynamics and shear relaxation of ionic liquids with long alkyl chain

The shear relaxation spectra of three imidazolium-based ionic liquids, 1-methyl-3-octylimidazolium chloride ([C(8)mim][Cl]), 1-methyl-3-octylimidazolium hexafluorophosphate ([C(8)mim][PF(6)]), and 1-dodecyl-3-methylimidazolium bis(trifluoromethanesulfonyl)amide ([C(12)mim][TFSA]) were measured and compared with the intermediate scattering functions determined with neutron spin echo (NSE) spectroscopy. The shear relaxation is slower than that predicted from the relaxation of the main peak of the structure factor that is common to other molecular liquids, whereas it is faster than that from the relaxation of the pre-peak, that corresponds to the correlation length of about 10 nm specific to ionic liquids with an intermediately long alkyl chain. The role of the pre-peak structure in the mechanism of shear viscosity of ionic liquids is discussed based on the comparison between NSE and shear relaxations.

Lipid Bilayers and Membrane Dynamics: Insight into Thickness Fluctuations

Thickness fluctuations have long been predicted in biological membranes but never directly observed experimentally. Here, we utilize neutron spin echo spectroscopy to experimentally reveal such fluctuations in a pure, fully saturated, phosphocholine lipid bilayer system. These fluctuations appear as an excess in the dynamics of undulation fluctuations. Like the bending rigidity, the thickness fluctuations change dramatically as the lipid transition temperature is crossed, appearing to be completely suppressed below the transition. Above the transition, the relaxation rate is on the order of 100 ns and is independent of temperature. The amplitude of the thickness fluctuations is 3.7 Å ± 0.7 Å, which agrees well with theoretical calculations and molecular dynamics simulations. The dependence of the fluctuations on lipid tail lengths is also investigated and determined to be minimal in the range of 14 to 18 carbon tails.

Scattering depth correction of evanescent waves in inelastic neutron scattering using a neutron prism

Grazing Incidence Neutron Spin Echo Spectroscopy (GINSES) has recently been applied to measure the dynamics of surfactant membranes close to a hydrophilic silicon wall. The scattering depth of the evanescent wave inside the microemulsion depends strongly on the angle of incidence and the wavelength. The inherently low scattering intensity of GINSES measurements, however, requires the integration over a rather broad wavelength band. In particular, at a pulsed source the instrument operates with a broad wavelength band covering all neutrons within one frame between two pulses. In order to yield viable counting statistics it is highly desirable to integrate data corresponding to significant fractions of the wavelength band. Therefore, in a normal reflectometry setup the penetration length would be smeared and blur the depth dependence of the experimental results. Here we describe a new method to strongly mitigate this effect and show its application in a GINSES experiment at the neutron spin echo instrument at the Spallation Neutron Source (SNS). A prism in front of the sample was introduced in order to adapt the angle of the incoming beam according to the wavelength by this optical component. As an example an experiment on a bicontinuous microemulsion using these neutron optics is presented.

Acceleration of membrane dynamics adjacent to a wall

The dynamics of an induced lamellar microemulsion adjacent to a planar hydrophilic surface (45 ns) were found to be three times faster compared to the bicontinuous bulk structure (133 ns). For these investigations the grazing incidence technique for neutron spin echo spectroscopy has been developed to resolve the depth dependent near surface dynamics. The observation is rationalized in terms of membrane hydrodynamics, where the flow fields reflected by the surface lead to a crossover from classical to confined fluctuations, and faster dynamics on large length scales (also known as "lubrication") are predicted.

Functional Domain Motions in Proteins on the ∼1–100 ns Timescale: Comparison of Neutron Spin-Echo Spectroscopy of Phosphoglycerate Kinase with Molecular-Dynamics Simulation

Protein function often requires large-scale domain motion. An exciting new development in the experimental characterization of domain motions in proteins is the application of neutron spin-echo spectroscopy (NSE). NSE directly probes coherent (i.e., pair correlated) scattering on the ∼1–100 ns timescale. Here, we report on all-atom molecular-dynamics (MD) simulation of a protein, phosphoglycerate kinase, from which we calculate small-angle neutron scattering (SANS) and NSE scattering properties. The simulation-derived and experimental-solution SANS results are in excellent agreement. The contributions of translational and rotational whole-molecule diffusion to the simulation-derived NSE and potential problems in their estimation are examined. Principal component analysis identifies types of domain motion that dominate the internal motion's contribution to the NSE signal, with the largest being classic hinge bending. The associated free-energy profiles are quasiharmonic and the frictional properties correspond to highly overdamped motion. The amplitudes of the motions derived by MD are smaller than those derived from the experimental analysis, and possible reasons for this difference are discussed. The MD results confirm that a significant component of the NSE arises from internal dynamics. They also demonstrate that the combination of NSE with MD is potentially useful for determining the forms, potentials of mean force, and time dependence of functional domain motions in proteins.

Microemulsions as model fluids for enhanced oil recovery: dynamics adjacent to planar hydrophilic walls

After the dynamics of microemulsions adjacent to a planar hydrophilic wall have been characterized using grazing incidence neutron spin echo spectroscopy, the model of Seifert was employed to explain the discovered acceleration for the surface near lamellar ordered membranes. Reflections of hydrodynamic waves by the wall - or the volume conservation between the membrane and the wall - explain faster relaxations and, therefore, a lubrication effect that is important for flow fields in narrow pores. The whole scenery is now spectated by using different scenarios of a bicontinuous microemulsion exposed to clay particles and of a lamellar microemulsion adjacent to a planar wall. The Seifert concept could successfully be transferred to the new problems.

Single Chain Dynamic Structure Factor of Poly(ethylene oxide) in Dynamically Asymmetric Blends with Poly(methyl methacrylate). Neutron Scattering and Molecular Dynamics Simulations

We have investigated the dynamically asymmetric polymer blend composed of short (Mn ≈ 2 kg/mol) poly(ethylene oxide) (PEO) and poly(methyl methacrylate) (PMMA) chains focusing on the collective dynamics of the fast PEO component. Using neutron spin-echo (NSE) spectroscopy, the single chain dynamic structure factor of PEO was investigated and compared to results from molecular dynamics simulations. After a successful validation of the simulations, a thorough analysis of the RPA approximation reveals the composition of the experimentally measured total scattering signal S(Q,t). Using the simulations, we show and calculate two contributions: (1) the relaxation of hydrogenated PEO against deuterated PEO, yielding the single chain dynamic structure factor of PEO, and (2) the relaxation of the PEO component against the PMMA matrix. For the short chains presented here the second contribution shows a significant decay at higher temperatures while it was previously shown that, in the case of long chains, no relaxation is found. This difference is related to a decrease of the glass transition temperature which takes place with decreasing chain length. In a second step we analyze the approximations that are used when calculating the single chain dynamic structure factor using the Rouse model. For a system like pure PEO, where the dynamics follow the predicted Rouse behavior, excellent agreement is achieved. In the case of PEO in PMMA, however, the slow PMMA matrix strongly influences the PEO dynamics. As a result, the distribution functions show a strong non-Gaussianity, and the calculation of S(Q,t) using the Rouse approximation fails even considering nonexponential Rouse mode correlators.

The intermediate scattering function for lipid bilayer membranes: From nanometers to microns

A numerical scheme based upon established hydrodynamic and elastic considerations is introduced and used to predict the intermediate scattering function for lipid bilayer membranes. The predictions span multiple wavelength regimes, including those studied by dynamic light scattering (DLS; microns) and neutron spin-echo (NSE) spectroscopy (10-100 nm). The results validate a recent theory specific to the NSE regime and expose slight inaccuracies associated with the theoretical results available in the DLS regime. The assumptions that underlie both our numerical methods and the related theoretical predictions are reviewed in detail to explain when certain results can be applied to experiment and where caution must be exercised.

Effect of charged lidocaine on static and dynamic properties of model bio-membranes

The effect of the charged lidocaine on the structure and dynamics of DMPC/DMPG (mass fraction of 95/5) unilamellar vesicles has been investigated. Changes in membrane organization caused by the presence of lidocaine were detected through small angle neutron scattering experiments. Our results suggest that the presence of lidocaine in the vicinity of the headgroups of lipid membranes leads to an increase of the area per lipid molecule and to a decrease of membrane thickness. Such changes in membrane structure may induce disordering of the tail group. This scenario explains the reduction of the main transition temperature of lipid membranes, as the fraction of lidocaine per lipid molecules increases, which was evident from differential scanning calorimetry results. Furthermore neutron spin echo spectroscopy was used for the dynamics measurements and the results reveal that presence of charged lidocaine increases the bending elasticity of the lipid membranes in the fluid phase and slows the temperature-dependent change of bending elasticity across the main transition temperature.

Light-Controlled Protein Dynamics Observed with Neutron Spin Echo Measurements

A photoresponsive surfactant has been used as a means to control protein structure and dynamics with light illumination. This cationic azobenzene surfactant, azoTAB, which undergoes a reversible photoisomerization upon exposure to the appropriate wavelength of light, adopts a relatively hydrophobic, trans structure under visible light illumination and a relatively hydrophilic cis structure under UV light illumination. Small-angle neutron scattering (SANS) and neutron spin echo (NSE) spectroscopy were used to measure the tertiary structure and internal dynamics of lysozyme in the presence of the photosurfactant, respectively. The SANS-based in vitro structures indicate that under visible light the photosurfactant induces partial unfolding that principally occurs away from the active site near the hinge region connecting the α and β domains. Upon UV exposure, however, the protein refolds to a nativelike structure. At the same time, enhanced internal dynamics of lysozyme were detected with the surfactant in the trans form through NSE measurements of the Q-dependent effective diffusion coefficient (D(eff)) of the protein. In contrast, the D(eff) values of lysozyme in the presence of cis azoTAB largely agree with the rigid-body calculation as well as those measured for pure lysozyme, suggesting that the native protein is dormant on the nanosecond time and nanometer length scales. Lysozyme internal motions were modeled by assuming a protein of two (α and β domains) or three (α and β domains and the hinge region) domains connects by either soft linkers or rigid, freely rotating bonds. Protein dynamics were also tracked with Fourier transform infrared spectroscopy through hydrogen-deuterium exchange kinetics, which further demonstrated enhanced protein flexibility induced by the trans form of the surfactant relative to the native protein. Ensemble-averaged intramolecular fluorescent resonance energy transfer measurements similarly demonstrated the enhanced dynamics of lysozyme with the trans form of the photosurfactant. Previous results have shown a significant increase in protein activity in the presence of azoTAB in the trans conformation. Combined, these results provide insight into a unique light-based method of controlling protein structure, dynamics, and function and strongly support the relevance of large domain motions for the activity of proteins.

Temperature and scattering contrast dependencies of thickness fluctuations in surfactant membranes

Temperature and scattering contrast dependencies of thickness fluctuations have been investigated using neutron spin echo spectroscopy in a swollen lamellar phase composed of nonionic surfactant, water, and oil. In the present study, two contrast conditions are examined; one is the bulk contrast, which probes two surfactant monolayers with an oil layer as a membrane, and the other is the film contrast, which emphasizes an individual surfactant monolayer. The thickness fluctuations enhance dynamics from the bending fluctuations, and are observed in a similar manner in both contrast conditions. Thickness fluctuations can be investigated regardless of the scattering contrast, though film contrasts are better to be employed in terms of the data quality. The thickness fluctuation amplitude is constant over the measured temperature range, including in the vicinity of the phase boundary between the lamellar and micellar phases at low temperature and the boundary between the lamellar and bicontinuous phases at high temperature. The damping frequency of the thickness fluctuations is well scaled using viscosity within the membranes at low temperature, which indicates the thickness fluctuations are predominantly controlled by the viscosity within the membrane. On the other hand, in the vicinity of the phase boundary at high temperature, thickness fluctuations become faster without changing the mode amplitude.

Nanoscale structures and dynamics of a boundary liquid layer

Our long term scientific interest is the understanding of the interface properties of flowingliquids on a microscopic level. Various mechanisms have been introduced to explain theorigin of slip at a solid–liquid interface like the formation of a thin depletion layeror a molecular ordering of the liquid near the interface. Reflectometry (usingx-rays or neutrons) is a powerful technique to probe structures in this surfaceregion. However, to date much less attention has been paid to the dynamicalproperties. In the first part of this paper we show that a different ordering of waterexists next to a hydrophobic substrate in comparison to a hydrophilic interface.Furthermore, we find that shear has no effect on the depletion layer on hydrophobicsubstrates, while no depletion layer exists for hydrophilic surfaces. The second partof the paper addresses the dynamical properties of the boundary layer, and wepresent a new method which enables the observation of the diffusion dynamicsof polymers next to a solid substrate. As a proof of concept, the dynamics ofmicelles next to the interface has been explored using grazing incidence neutronspin-echo spectroscopy. We were able to verify that investigation of the dynamicsof the sample is feasible with this grazing incidence technique and we presentdata taken near the critical angle of total reflection. It appears that the diffusivemotion of micelles at the hydrophobic (repulsive) interface is faster than at ahydrophilic interface or in the bulk. Furthermore, neutron spin-echo spectroscopywas extended to a first evaluation of the Doppler shift which occurs under flow.