Neutron Spin Echo Measurements Research Articles

From ionic clusters dynamics to network constraints in ionic polymer solutions.

Physical networks formed by ionizable polymers with ionic clusters as crosslinks are controlled by coupled dynamics that transcend from ionic clusters through chain motion to macroscopic response. Here, the coupled dynamics, across length scales, from the ionic clusters to the networks in toluene swollen polystyrene sulfonate networks, were directly correlated, as the electrostatic environment of the physical crosslinks was altered. The multiscale insight is attained by coupling neutron spin echo measurements with molecular dynamics simulations, carried out to times typical of relaxation of polymers in solutions. The experimental dynamic structure factor is in outstanding agreement with the one calculated from computer simulations, as the networks are perturbed by elevating the temperature and changing the electrostatic environment. In toluene, the long-lived clusters remain stable over hundreds of ns across a broad temperature range, while the polymer network remains dynamic. Though the size of the clusters changes as the dielectric constant of the solvent is modified through the addition of ethanol, they remain stable but morph, enhancing the polymer chain dynamics.

Internal dynamics of multidomain protein as revealed by an optimized neutron spin echo measurement and all-atom molecular dynamics simulation

Internal dynamics of multidomain protein as revealed by an optimized neutron spin echo measurement and all-atom molecular dynamics simulation

Impact of transmembrane peptides on individual lipid motions and collective dynamics of lipid bilayers

The fluid nature of lipid bilayers is indispensable for the dynamic regulation of protein function and membrane morphology in biological membranes. Membrane-spanning domains of proteins interact with surrounding lipids and alter the physical properties of lipid bilayers. However, there is no comprehensive view of the effects of transmembrane proteins on the membrane's physical properties. Here, we investigated the effects of transmembrane peptides with different flip-flop-promoting abilities on the dynamics of a lipid bilayer employing complemental fluorescence and neutron scattering techniques. The quasi-elastic neutron scattering and fluorescence experiments revealed that lateral diffusion of the lipid molecules and the acyl chain motions were inhibited by the inclusion of transmembrane peptides. The neutron spin-echo spectroscopy measurements indicated that the lipid bilayer became more rigid but more compressible and the membrane viscosity increased when the transmembrane peptides were incorporated into the membrane. These results suggest that the inclusion of rigid transmembrane structures hinders individual and collective lipid motions by slowing down lipid diffusion and increasing interleaflet coupling. The present study provides a clue for understanding how the local interactions between lipids and proteins change the collective dynamics of the lipid bilayers, and therefore, the function of biological membranes.

Molecular simulations and NMR reveal how lipid fluctuations affect membrane mechanics.

The lipid make-up of a bilayer determines its measurable properties but how the motions of individual molecules combine to produce these properties remains unclear. By exploiting the synergy between NMR spectroscopy and molecular dynamics (MD) simulations, we show that the lipid dynamics in a bilayer are collective yet segmental in nature and contribute directly to bilayer elasticity. Our analysis entails development of a theoretical framework that allows direct comparison of carbon-hydrogen (C-H) bond relaxations as measured by simulations and solid-state NMR experiments. The new formalism allows validation of lipid force fields against NMR data by considering a fixed bilayer normal (director axis) and restricted anisotropic motion of the C-H bonds described by segmental order parameters. The simulations showed that the spin-lattice relaxation rates scale as the squared order parameter of the lipid acyl chains, yielding an apparent bending modulus kc of the bilayer. Our results mirror the experimental observations of kc obtained from solid-state NMR studies of 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC) bilayers with increasing amounts of cholesterol. These findings are further validated by neutron spin-echo measurements of membrane bending fluctuations and emergent elastic properties. Analysis of the thermally excited C-H bond fluctuations from the simulations also reveals collective dynamics of a nematic-like nature and uncovers a critical role of interleaflet coupling in membrane mechanics. This study thus provides novel insights into the inner workings of lipid membranes and establishes a new tool for validating computational approaches against experimental data.

Molecular simulations and NMR reveal how lipid fluctuations affect membrane mechanics

Lipid bilayers form the main matrix of functional cell membranes, and their dynamics underlie a host of physical and biological processes. Here we show that elastic membrane properties and collective molecular dynamics (MD) are related by the mean-square amplitudes (order parameters) and relaxation rates (correlation times) of lipid acyl chain motions. We performed all-atom MD simulations of liquid-crystalline bilayers that allow direct comparison with carbon-hydrogen (CH) bond relaxations measured with NMR spectroscopy. Previous computational and theoretical approaches have assumed isotropic relaxation, which yields inaccurate description of lipid chain dynamics and incorrect data interpretation. Instead, the new framework includes a fixed bilayer normal (director axis) and restricted anisotropic motion of the CH bonds in accord with their segmental order parameters, enabling robust validation of lipid force fields. Simulated spectral densities of thermally excited CH bond fluctuations exhibited well-defined spin-lattice (Zeeman) relaxations analogous to those in NMR measurements. Their frequency signature could be fit to a simple power-law function, indicative of nematic-like collective dynamics. Moreover, calculated relaxation rates scaled as the squared order parameters yielding an apparent κC modulus for bilayer bending. Our results show a strong correlation with κC values obtained from solid-state NMR studies of bilayers without and with cholesterol as validated by neutron spin-echo measurements of membrane elasticity. The simulations uncover a critical role of interleaflet coupling in membrane mechanics and thus provide important insights into molecular sites of emerging elastic properties within lipid bilayers.

Quantifying structure and dynamics of bound and bulk polymer in tailor-made rubber-silica nanocomposites

The dynamics of long polymer chains in the presence of nanoparticles have been investigated. The nanocomposites of interest were inspired by tire industry-like rubber materials and consisted of entangled polyisoprene linear chains mixed with chemically pre-treated silica nanoparticles. Combining rheology, dielectric spectroscopy, and neutron spin echo measurements, we measured the modification of the polymer chain dynamics from bulk state to high filler concentration over a broad range of time and length scales. We show that the end-to-end relaxation does not seem to be impacted, whereas the polymer dynamics is significantly slowed down at a very local scale in the presence of nano-fillers. In addition to this length scale dependent different dynamics, additional Neutron Spin Echo spectroscopy experiments and Small Angle Scattering on labelled polymer chains, irreversibly bound to the filler surface and re-dispersed in a fully deuterated matrix, revealed a negligible dynamical behavior of this particular population of localized chains in the rubber layer. The conformational statistics of these chains is that of self-avoiding walk train within a shell that is thinner than the size of the chain. To the best of our knowledge, this work is the first of its kind which measures the single chain form factor in the bound layer of chemically multi-linked chains to the filler surface.

Topological Dynamics of Micelles Formed by Geometrically Varied Surfactants

The molecular architecture of sugar-based surfactants strongly affects their self-assembled structure, i.e., the type of micelles they form, which in turn controls both the dynamics and rheological properties of the system. Here, we report the segmental and mesoscopic structure and dynamics of a series of C16 maltosides with differences in the anomeric configuration and degree of tail unsaturation. Neutron spin-echo measurements showed that the segmental dynamics can be modeled as a one-dimensional array of segments where the dynamics increase with inefficient monomer packing. The network dynamics as characterized by dynamic light scattering show different relaxation modes that can be associated with the micelle structure. Hindered dynamics are observed for arrested networks of worm-like micelles, connected to their shear-thinning rheology, while nonentangled diffusing rods relate to Newtonian rheological behavior. While the design of novel surfactants with controlled properties poses a challenge for synthetic chemistry, we demonstrate how simple variations in the monomer structure can significantly influence the behavior of surfactants.

Viscosity reduction in polymer nanocomposites: Insights from dynamic neutron and X‐ray scattering

AbstractThe addition of nanoparticles to a polymer matrix can in certain cases induce a reduction in viscosity, with respect to the pure matrix, in the resulting composites. This counterintuitive phenomenon cannot be explained using the most common rheological models. For this reason, it has been chosen as a good example in this paper to demonstrate the value and methods of dynamic X‐ray and neutron scattering techniques for the investigation of polymer nanocomposites. An overview of the main results on this topic is presented together with an introduction to the basic concepts relating to X‐ray photon correlation spectroscopy, neutron backscattering, and neutron spin echo measurements.

Pluronic-based lamellar phases: influence of polymer architecture on bilayer bending elasticity

ABSTRACT In this study, we investigate the influence of the block copolymer architecture on the bending elasticity of polymer-rich lamellar phases. In detail, we study the polymer-rich system water/o-xylene/Pluronic triblock copolymer/CTAB and change the polymer while keeping the mass ratios constant. We use neutron spin echo measurements (NSE) to determine κ. With the measurements we determine the relaxation rate of the bilayer bending mode and subsequently calculate the bilayer bending modulus κ using the Zilman-Granek approach [Zilman et al., Phys. Rev. Lett. 77, 4788–4791 (1996)]. Moreover, we have investigated the lamellar phases via small-angle X-ray scattering (SAXS) and have analysed the data with the modified Caillé theory to obtain the Caillé parameter η. Based on κ the bulk compression modulus can be calculated. Increasing values of κ are found with increasing EO and PO block size in the range of 3.8 to 13.4 T. The computed values of are in the range of 10 to 10 Pa.

Data Analysis and Background Subtraction in Neutron Spin Echo Spectroscopy

With the constantly improving performance of neutron spin echo (NSE) spectrometers it becomes possible to perform measurements on increasingly complex samples and to study more and more delicate effects. To properly study such effects, proper background correction becomes increasingly important. In this paper, we will review different methods to subtract the buffer from NSE measurements and study the effect of small errors in the subtraction of the background. In the large dynamic range of modern neutron spin-echo spectrometers multiple effects become visible in a single measurement. Specifically, for vesicles both membrane undulations and translational diffusion have an effect on the intermediate scattering function in the NSE time window and here, we will investigate how taking this into account differently affects the results obtained from data analysis.

Using Molecular Dynamics to Gain Atomic-Level Insight into Bilayer Properties Measured with NMR and Neutron Spin-Echo Spectroscopy

Lipid bilayers form the core of functional membrane platforms such as cellular plasma membranes. Their dynamics are inherently related to a wide range of physical and biological processes and thus have become the subject of both experimental measurements and theoretical investigations. Nuclear magnetic resonance (NMR) in particular, is able to probe a hierarchy of lipid motions by measuring the power spectrum of thermally excited molecular fluctuations. From this spectrum of molecular noise emerge both average conformational parameters such as chain order, SCD, and dynamical descriptors such as the relaxation rate, R1, of carbon-hydrogen (or deuterium) bond fluctuations. Interestingly, these experimental observables exhibit specific functional relationships including an SCD-R1 square-law dependence, and an R1 inverse square-root frequency law. Different theories explain these functional forms, but the convoluted nature of the NMR data complicates further decoupling of the signal and testing of underlying models. Here we use atomistic molecular dynamics (MD) simulations to investigate the underpinnings of the experimental results. We calculated SCD and R1 for a series of experimentally studied bilayers composed of unsaturated or saturated lipids with different cholesterol content and at different hydration levels. The simulation results show the same functional dependencies as those observed with NMR and reveal unprecedented details of the relevant molecular mechanisms. Following a recently developed paradigm, we use the slope of the SCD-R1 square-law dependence from the simulations to calculate the bilayer bending rigidity modulus kC and explore its underlying parameter space. Comparison to kC values obtained from MD splay fluctuations and neutron spin-echo measurements allows for a closer look at the complementary applications of the techniques. Our results provide novel and clarifying insights into the nature of experimentally accessible molecular motions and their biophysical implications.

Membrane Deformation Modulated by Hydration and Cholesterol Unveiled by Solid-State 2H NMR Spectroscopy

Phospholipid membranes are biological liquid crystals in which intermolecular interactions at mesoscopic length scales play key roles in the emergence of membrane physical properties like structure and rigidity. Such interactions modulated by membrane hydration [1] and cholesterol [2] have been investigated using solid-state 2H NMR spectroscopy. The average structure is manifested by the lineshapes through principal values of static or residual coupling tensors due to quadrupolar interactions. The residual quadrupolar couplings (RQCs) yield segmental order parameters for the individual acyl segments (i), providing average membrane properties such as bilayer thickness, area/lipid, and spontaneous curvature [3]. We show that membrane dehydration and increased cholesterol content result in greater RQCs, indicative of membrane thickening. Elastic membrane deformations are exhibited as collective order-director fluctuations (ODF) contributing to the nuclear magnetic relaxation. At mesoscopic length scales the thermal fluctuations connect to membrane mechanical properties via the fluctuation-dissipation theorem. Model-free interpretation of the functional dependence of spin-lattice relaxation rates, R1Z, on segmental order parameters (square-law) reveals emergent material properties in the liquid-crystalline state. The R1Z relaxation dispersion and temperature dependence of square-law plots indicate a nematic-like membrane hydrocarbon core. Solid-state 2H NMR relaxation studies demonstrate that both dehydration and increased cholesterol content in the membrane lead to decreased slopes of square-law plots, indicating an increase in membrane bending rigidity, consistent with neutron spin-echo (NSE) measurements. Analogous 2H NMR studies with various membrane lipid compositions and peptide-reconstituted membranes have addressed lipid-mediated cell signaling and lipid-protein interactions. Molecular dynamics simulations provide further insights on the correspondence of the solid-state NMR and NSE results. [1] K.J. Mallikarjunaiah et al. (2019) Phys. Chem. Chem. Phys. 21, 18422. [2] S. Chakraborty et al. (2020) Proc. Natl. Acad. Sci. 117, 21896. [3] Mallikarjunaiah et al. (2011) Biophys. J. 100, 98.

In Situ Dynamic Light Scattering Complementing Neutron Spin Echo Measurements on Protein Samples

Monitoring the state of the sample on the minute-time scale is crucial in case of sensitive soft matter or biological samples, given that neutron spin echo measurements take up to several days. Moreover, there is no method to interpret the normalized intermediate scattering function obtained by measuring the neutron spin echo if relevant sample properties change during measurement. Dynamic light scattering provides information on the diffusion constant of particles in solution (biological macromolecules, such as proteins, protein aggregates, polymer particles, etc.) with average hydrodynamic radii in a wide range from several nanometers up to several microns. This information can be obtained in a few minutes and it offers a good overview of the current sample state. Details on the novel in situ dynamic light scattering set-up with one fixed scattering angle and first results obtained on a molten globule state of apo-myoglobin are presented.

Revealing the Aggregation Mechanism, Structure, and Internal Dynamics of Poly(vinyl alcohol) Microgel Prepared through Liquid–Liquid Phase Separation

The use of technologies based on soft polymer particles represents an effective way to deliver target molecules with a specific function. To design a well-performing delivery system, it is fundamental to rationalize both the aggregation and the structural properties of such particles. In this study, we present the kinetic and structural characterization over time of poly(vinyl alcohol) (PVA) microgels obtained through a salting-out process in the presence of NaCl. We have analyzed how both the polymer and salt concentrations affect the aggregation process. The aggregation rate as well as the morphology and physico-chemical parameters, such as mass and chain density of the microgels, have been determined through static and dynamic light scattering and discussed in the framework of the diffusion-limited and reaction-limited colloid aggregation. Insights into the polymer chain arrangements and their dynamics have been gained by means of small-angle neutron scattering and neutron spin-echo measurements. As a ...

Dynamics in polyelectrolyte/microemulsion complexes

Oil-in-water (O/W) microemulsion droplets are convenient carriers for hydrophobic molecules in an aqueous phase and are used for a wide range of applications. We studied weakly charged O/W microemulsion droplets complexed with oppositely charged polyacrylates that form long linear arrangements of droplets. All samples showed rather low viscosities, which is in contrast to similar systems of hydrophobically interconnected droplets. Here, we applied small-angle neutron scattering, dynamic light scattering and neutron spin-echo spectroscopy to characterise the dynamic properties of polyacrylate/microemulsion complexes in order to understand the origin of the low-viscous behaviour. We found that the electrostatic interactions lead to very dynamic complexes with high exchange rates of droplets and only a fraction of the droplets is contained within the transient complexes at a given time. These results were only accessible by the combination of different methods as one method alone would have given an incomplete picture.

Direct measurement of topological interactions in polymers under shear using neutron spin echo spectroscopy

We present in-situ neutron spin echo measurements on an entangled polydimethylsiloxane melt under shear and demonstrate the ability to monitor nano-scale dynamics in flowing liquids. We report no changes in the topological interactions of the chains for shear rates approaching the inverse longest relaxation time. Further experiments following along this line will allow to systematically test the predictions of theories, like e.g. convective constraint release.

Detecting Branching in Wormlike Micelles via Dynamic Scattering Methods.

Topological differences in a model series of wormlike micelles (WLMs) with varying degrees of branching are investigated via neutron spin echo (NSE) and dynamic light scattering (DLS). Branching was previously verified via microscopy, scattering methods, and rheology. NSE measurements reveal that on the shortest length scales, where WLMs are topologically similar, the dynamic behavior is independent of branching level. However, branching leads to deviations from wormlike chain behavior on length scales relevant to branching. These differences continue into the DLS regime, where two relaxation modes are observed. While measuring micellar branching is challenging, our measurements of segmental dynamics provide a direct dynamic measurement of WLM branching and demonstrate promise for future identification of topological changes in self-assembled materials.

Short-time dynamics of lysozyme solutions with competing short-range attraction and long-range repulsion: Experiment and theory.

Recently, atypical static features of microstructural ordering in low-salinity lysozyme protein solutions have been extensively explored experimentally and explained theoretically based on a short-range attractive plus long-range repulsive (SALR) interaction potential. However, the protein dynamics and the relationship to the atypical SALR structure remain to be demonstrated. Here, the applicability of semi-analytic theoretical methods predicting diffusion properties and viscosity in isotropic particle suspensions to low-salinity lysozyme protein solutions is tested. Using the interaction potential parameters previously obtained from static structure factor measurements, our results of Monte Carlo simulations representing seven experimental lysoyzme samples indicate that they exist either in dispersed fluid or random percolated states. The self-consistent Zerah-Hansen scheme is used to describe the static structure factor, S(q), which is the input to our calculation schemes for the short-time hydrodynamic function, H(q), and the zero-frequency viscosity η. The schemes account for hydrodynamic interactions included on an approximate level. Theoretical predictions for H(q) as a function of the wavenumber q quantitatively agree with experimental results at small protein concentrations obtained using neutron spin echo measurements. At higher concentrations, qualitative agreement is preserved although the calculated hydrodynamic functions are overestimated. We attribute the differences for higher concentrations and lower temperatures to translational-rotational diffusion coupling induced by the shape and interaction anisotropy of particles and clusters, patchiness of the lysozyme particle surfaces, and the intra-cluster dynamics, features not included in our simple globular particle model. The theoretical results for the solution viscosity, η, are in qualitative agreement with our experimental data even at higher concentrations. We demonstrate that semi-quantitative predictions of diffusion properties and viscosity of solutions of globular proteins are possible given only the equilibrium structure factor of proteins. Furthermore, we explore the effects of changing the attraction strength on H(q) and η.

Modulation of hemoglobin dynamics by an allosteric effector.

Hemoglobin (Hb) is an extensively studied paradigm of proteins that alter their function in response to allosteric effectors. Models of its action have been used as prototypes for structure‐function relationships in many proteins, and models for the molecular basis of its function have been deeply studied and extensively argued. Recent reports suggest that dynamics may play an important role in its function. Relatively little is known about the slow, correlated motions of hemoglobin subunits in various structural states because experimental and computational strategies for their characterization are challenging. Allosteric effectors such as inositol hexaphosphate (IHP) bind to both deoxy‐Hb and HbCO, albeit at different sites, leading to a lowered oxygen affinity. The manner in which these effectors impact oxygen binding is unclear and may involve changes in structure, dynamics or both. Here we use neutron spin echo measurements accompanied by wide‐angle X‐ray scattering to show that binding of IHP to HbCO results in an increase in the rate of coordinated motions of Hb subunits relative to one another with little if any change in large scale structure. This increase of large‐scale dynamics seems to be coupled with a decrease in the average magnitude of higher frequency modes of individual residues. These observations indicate that enhanced dynamic motions contribute to the functional changes induced by IHP and suggest that they may be responsible for the lowered oxygen affinity triggered by these effectors.

A neutron spin echo study of low-temperature water confined in the spherical silica pores of SBA-16.

The dynamic properties of heavy water (D2O) and light water (H2O) confined in porous silica SBA-16 were studied over a temperature range of 210-290 K by neutron spin echo measurements. SBA-16 has predominant spherical pores (7.1 nm in pore size), channels interconnecting the spherical pores, and micropores (corona). The coherent intermediate scattering function on D2O filled SBA-16 showed the rotational dynamics of confined water without significant translational motion over the temperature range measured. This finding is due probably to collective entities of water due to cooperativity of hydrogen-bonds among water molecules in SBA-16 pores. The relaxation time of the collective entities followed the Vogel-Fulcher-Tammann relation at temperatures down to the freezing temperature of 235 K, suggesting a behavior of fragile water in the spherical pore. A comparison with previous NSE measurements of D2O in MCM-41 showed that the collective entities of water in the SBA-16 spherical pores have higher rotational mobility than those in the MCM-41 cylindrical pores. On the other hand, the incoherent intermediate scattering function on H2O filled SBA-16 revealed the translational motion of individual water molecules in the collective entities. It has been found that water in micropores is not frozen and is mobile down to 210 K from data of both D2O and H2O in SBA-16. Phase changes of various water confined in SBA-16 with decreasing and increasing temperatures are discussed based on the obtained dynamic properties.