Neutron Spin Echo Research Articles

A Needless but Interesting Controversy with Pedagogical Overtones

A decade ago four independent groups, using four independent methods, reported the unanticipated and still surprising result that the bending modulus KC of bilayers of DOPC, unlike other standard lipids, does not increase with addition of cholesterol. A recent paper, using neutron spin echo (NSE) and nuclear magnetic resonance relaxation times (NMRτ), reports that the bending modulus of DOPC increases three fold with increasing cholesterol concentration up to 50%. It would appear that this has become a controversial topic. This controversy is needless because NSE and NMRτ do not measure the bending modulus as it has been defined for decades. Each measures important, complementary, properties that could enrich our understanding of bilayer properties, notably membrane viscosity and location of the neutral plane employed in membrane mechanics. The resolution of this controversy rests on elucidating the difference between measurements on a system in equilibrium that lead directly to equilibrium properties like the bending modulus, and measurements on the same equilibrium system that lead to more complex results involving both static moduli and transport properties. While the difference between static and non-equilibrium quantities is well understood in the physical sciences and statistical thermodynamics, this emerging controversy seems to warrant a pedagogical reminder.

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.

Melittin exerts opposing effects on short- and long-range dynamics in bicontinuous microemulsions

Bicontinuous microemulsions (BμEs) are a promising biomembrane mimetic system for investigating the behavior of antimicrobial peptides (AMPs) and their delivery to open wounds to combat antibiotic-resistant microorganisms. The properties of the BμE host are in turn affected by the guest AMP and can deviate from those of the unperturbed BμEs, especially at higher AMP concentrations. Here we report the effect of an archetypal AMP, melittin, over a wide range of concentrations, on the nanoscopic dynamics of BμEs formed by water/sodium dodecyl sulfate (SDS)/1-pentanol/dodecane, investigated using quasi-elastic neutron scattering (QENS). Two distinct motions are observed, namely, (i) the lateral motion of the surfactant on the surface of the oil channels and (ii) the internal motion of the surfactants. It is found that melittin restricts both the lateral and the internal motion, thereby acting as a stiffening agent. The lateral motion is more strongly affected, at low concentration of melittin. The lateral diffusion coefficient decreased sharply, approaching a constant value at higher melittin concentration. These results are in sharp contrast with the recent dynamic light scattering and neutron spin echo results which showed that at the length and time scales longer than those probed in the current work, melittin enhanced the long-range collective and local undulation motions of BμEs. Considered together, our results indicate that incorporation of melittin modulates the dynamics differently depending on the spatial and temporal regimes, in which the dynamics are being probed. The addition of melittin at low concentrations increased the magnitude of the zeta potential, but further increase of the melittin concentration decreased it. This suggests that addition of melittin at low concentrations led to increase in the surfactant concentration, but did not affect the negative charge per surfactant molecule, while further addition of melittin led to ion pairing of melittin with the oppositely charged surfactant. This study therefore demonstrates how the addition of melittin hinders the lateral motion of surfactants as a result of the strong association between melittin and SDS, suggesting that the release of AMPs from BμE-based delivery vehicles may be hindered.

Evolution of helimagnetic correlations when approaching the quantum critical point of Mn1−xFexSi

We present a comprehensive investigation of the evolution of helimagnetic correlations in Mn$_{1-x}$Fe$_x$Si with increasing doping. By combining polarised neutron scattering and high resolution Neutron Spin Echo spectroscopy we investigate three samples with $x$=0.09, 0.11 and 0.14, i.e. with compositions on both sides of the concentration $x^* \sim 0.11$ where the helimagnetic Bragg peaks disappear and between $x^*$ and the quantum critical concentration $x_C \sim 0.17$, where $T_C$ vanishes. We find that the abrupt disappearance of the long range helical periodicity at $x^*$, does not affect the precursor fluctuating correlations. These build up with decreasing temperature in a similar way as for the parent compound MnSi. Also the dynamics bears strong similarities to MnSi. The analysis of our results indicates that frustration, possibly due to achiral RKKY interactions, increases with increasing Fe doping. We argue that this effect explains both the expansion of the precursor phase with increasing $x$ and the abrupt disappearance of long range helimagnetic periodicity at $x^*$.

Surfactant Monolayer Bending Elasticity in Lipase Containing Bicontinuous Microemulsions.

Lipase-catalyzed reactions offer many advantages among which a high degree of selectivity combined with the possibility to convert even non-natural substrates are of particular interest. A major drawback in the applicability of lipases in the conversion of synthetically interesting, non-natural substrates is the substantial insolubility of such substrates in water. The conversion of substrates, natural or non-natural, by lipases generally involves the presence of a water–oil interface. In the present paper, we exploit the fact that the presence of lipases, in particular the lipase from Candida antarctica B (CalB), changes the bending elastic properties of a surfactant monolayer in a bicontinuous microemulsion consisting of D2O/NaCl -n-(d)-octane-pentaethylene glycol monodecyl ether (C10E5) in a similar manner as previously observed for amphiphilic block-copolymers. To determine the bending elastic constant, we have used two approaches, small angle neutron scattering (SANS) and neutron spin echo (NSE) spectroscopy. The time-averaged structure from SANS showed a slight decrease in bending elasticity, while on nanosecond time scales as probed with NSE, a stiffening has been observed, which was attributed to adsorption/desorption mechanisms of CalB at the surfactant monolayer. The results allow to derive further information on the influence of CalB on the composition and bending elasticity of the surfactant monolayer itself as well as the underlying adsorption/desorption mechanism.

Molecular spin echoes; multiple magnetic coherences in molecule surface scattering experiments.

In this paper we demonstrate that a molecular beam of hydrogen molecules can be magnetically manipulated to produce multiple coherences in the molecular interference pattern. Unlike spin 1/2 magnetic beam experiments, i.e., neutron and helium spin echo, the nuclear and rotational magnetic moments in a molecule are strongly coupled. We show experimentally and theoretically that this coupling leads to multiple magnetic field conditions under which the magnetic moment of molecules travelling with different speeds can be coherently refocussed. We also demonstrate that these multiple coherence signals are extremely sensitive to the scattering event, opening up new possibilities for measuring molecule-surface interactions.

Collective dynamics in lipid membranes containing transmembrane peptides.

Biological membranes are composed of complex mixtures of lipids and proteins that influence each other's structure and function. The biological activities of many channel-forming peptides and proteins are known to depend on the material properties of the surrounding lipid bilayer. However, less is known about how membrane-spanning channels affect the lipid bilayer properties, and in particular, their collective fluctuation dynamics. Here we use neutron spin echo spectroscopy (NSE) to measure the collective bending and thickness fluctuation dynamics in dimyristoylphosphatidylcholine (di 14 : 0 PC, DMPC) lipid membranes containing two different antimicrobial peptides, alamethicin (Ala) and gramicidin (gD). Ala and gD are both well-studied antimicrobial peptides that form oligomeric membrane-spanning channels with different structures. At low concentrations, the peptides did not have a measurable effect on the average bilayer structure, yet significantly changed the collective membrane dynamics. Despite both peptides forming transmembrane channels, they had opposite effects on the relaxation time of the collective bending fluctuations and associated effective bending modulus, where gD addition stiffened the membrane while Ala addition softened the membrane. Meanwhile, the lowest gD concentrations enhanced the collective thickness fluctuation dynamics, while the higher gD concentrations and all studied Ala concentrations dampened these dynamics. The results highlight the synergy between lipids and proteins in determining the collective membrane dynamics and that not all peptides can be universally treated as rigid bodies when considering their effects on the lipid bilayer fluctuations.

Collective Motions and Mechanical Response of a Bulk of Single-Chain Nano-Particles Synthesized by Click-Chemistry.

We investigate the effect of intra-molecular cross-links on the properties of polymer bulks. To do this, we apply a combination of thermal, rheological, diffraction, and neutron spin echo experiments covering the inter-molecular as well as the intermediate length scales to melts of single-chain nano-particles (SCNPs) obtained through ‘click’ chemistry. The comparison with the results obtained in a bulk of the corresponding linear precursor chains (prior to intra-molecular reaction) and in a bulk of SCNPs obtained through azide photodecomposition process shows that internal cross-links do not influence the average inter-molecular distances in the melt, but have a profound impact at intermediate length scales. This manifests in the structure, through the emergence of heterogeneities at nanometric scale, and also in the dynamics, leading to a more complex relaxation behavior including processes that allow relaxation of the internal domains. The influence of the nature of the internal bonds is reflected in the structural relaxation that is slowed down if bulky cross-linking agents are used. We also found that any residual amount of cross-links is critical for the rheological behavior, which can vary from an almost entanglement-free polymer bulk to a gel. The presence of such inter-molecular cross-links additionally hinders the decay of density fluctuations at intermediate length scales.

Neutron Spin–Echo Studies of the Structural Relaxation of Network Liquid ZnCl2 at the Structure Factor Primary Peak and Prepeak

Using neutron spin-echo spectroscopy, we studied the microscopic structural relaxation of a prototypical network ionic liquid ZnCl2 at the structure factor primary peak and prepeak. The results show that the relaxation at the primary peak is faster than the prepeak and that the activation energy is ∼33% higher. A stretched exponential relaxation is observed even at temperatures well-above the melting point Tm. Surprisingly, the stretching exponent shows a rapid increase upon cooling, especially at the primary peak, where it changes from a stretched exponential to a simple exponential on approaching the Tm. These results suggest that the appearance of glassy dynamics typical of the supercooled state even in the equilibrium liquid state of ZnCl2 as well as the difference of activation energy at the two investigated length scales are related to the formation of a network structure on cooling.

Crystallization of magnetic skyrmions in MnSi investigated by neutron spin echo spectroscopy

The authors study crystallization process of magnetic skyrmions in a chiral magnet MnSi by means of a resonance-type neutron spin echo spectroscopy in a magnetic field, and find that the skyrmions aggregate into small static triangular-lattice domains near the critical temperature.

Bridging dynamic regimes of segmental relaxation and center-of-mass diffusion in associative protein hydrogels

Molecular transport in dense, crowded media is often affected by binding and collisions with obstacles, leading to complex spatial distributions and anomalous diffusion on multiple length scales. Here, neutron spin-echo (NSE) spectroscopy and forced Rayleigh scattering (FRS) were used in tandem to study the interplay between segmental and center-of-mass chain dynamics in the presence of strong binding interactions in a model protein hydrogel. The results provide evidence for several dynamic scaling regimes of gel relaxation behavior with varying length scale, including a caging regime bridging submolecular relaxation and center-of-mass diffusion due to transient binding. On mesoscopic length scales larger than the size of the cage, chains undergo two distinct regimes of apparent superdiffusive scaling, with different power-law exponents, before the onset of terminal Fickian diffusion. The combined NSE and FRS results are interpreted in the context of prior Brownian dynamics simulations of associating star polymers, revealing insight into structural length scales and binding kinetics governing the transition from segmental relaxation to self-diffusion in the protein hydrogel. Finally, single-sticker tracer diffusion measurements were performed to directly probe sticker association/dissociation dynamics within the gel, the results suggesting that cooperative cluster motion may play a role in network relaxation on larger length scales.Received 28 February 2020Accepted 30 October 2020DOI:https://doi.org/10.1103/PhysRevResearch.2.043369Published by the American Physical Society under the terms of the Creative Commons Attribution 4.0 International license. Further distribution of this work must maintain attribution to the author(s) and the published article's title, journal citation, and DOI.Published by the American Physical SocietyPhysics Subject Headings (PhySH)Research AreasDiffusionPhysical SystemsPolymer gelsTechniquesLight scatteringNeutron scatteringNeutron spin echo spectroscopyPolymers & Soft Matter

Understanding the Structure and Dynamics of Complex Biomembrane Interactions by Neutron Scattering Techniques.

The membrane is one of the key structural materials of biology at the cellular level. Composed predominantly of a bilayer of lipids with embedded and bound proteins, it defines the boundaries of the cell and many organelles essential to life and therefore is involved in almost all biological processes. Membrane-specific interactions, such as drug binding to a membrane receptor or the interactions of an antimicrobial compound with the lipid matrix of a pathogen membrane, are of interest across the scientific disciplines. Herein we present a review, aimed at nonexperts, of the major neutron scattering techniques used in membrane studies: small-angle neutron scattering, neutron membrane diffraction, neutron reflectometry, quasielastic neutron scattering, and neutron spin echo. Neutron scattering techniques are well suited to studying biological membranes. The nondestructive nature of cold neutrons means that samples can be measured for long periods without fear of beam damage from ultraviolet, electron, or X-ray radiation, and neutron beams are highly penetrating, thus offering flexibility in samples and sample environments. Most important is the strong difference in neutron scattering lengths between the two most abundant forms of hydrogen, protium and deuterium. Changing the relative amounts of protium/deuterium in a sample allows the production of a series of neutron scattering data sets, enabling the observation of differing components within complex membrane architectures. This approach can be as simple as using the naturally occurring neutron contrast between different biomolecules to study components in a complex by changing the solution H2O/D2O ratio or as complex as selectively labeling individual components with hydrogen isotopes. This review presents an overview of each experimental technique with the neutron instrument configuration, related sample preparation and sample environment, and data analysis, highlighted by a special emphasis on using prominent neutron contrast to understand structure and dynamics. This review gives researchers a practical introduction to the often enigmatic suite of neutron beamlines, thereby lowering the barrier to taking advantage of these large-facility techniques to achieve new understandings of membranes and their interactions with other molecules.

Self-Similar Dynamics of Large Polymer Rings: A Neutron Spin Echo Study.

This work clarifies the self-similar dynamics of large polymer rings using pulsed-field gradient nuclear magnetic resonance and neutron spin echo spectroscopy. We find center of mass diffusion taking place in three dynamic regimes starting (i)with a strongly subdiffusive domain ⟨r^{2}(t)⟩_{com}∼t^{α} (0.4≤α≤0.65); (ii)a second subdiffusive region ⟨r^{2}(t)⟩_{com}∼t^{0.75} that (iii)finally crosses over to Fickian diffusion. While the t^{0.75} range previously has been found in simulations and was predicted by theory, we attribute the first to the effect of cooperative dynamics resulting from the correlation hole potential. The internal dynamics at scales below the elementary loop size is well described by ring Rouse motion. At larger scales the dynamics is self-similar and follows very well the predictions of the scaling models with preference for the self-consistent fractal loopy globule model.

Crowding in the Eye Lens: Modeling the Multisubunit Protein β-Crystallin with a Colloidal Approach

We present a multiscale characterization of aqueous solutions of the bovine eye lens protein βH crystallin from dilute conditions up to dynamical arrest, combining dynamic light scattering, small-angle x-ray scattering, tracer-based microrheology, and neutron spin echo spectroscopy. We obtain a comprehensive explanation of the observed experimental signatures from a model of polydisperse hard spheres with additional weak attraction. In particular, the model predictions quantitatively describe the multiscale dynamical results from microscopic nanometer cage diffusion over mesoscopic micrometer gradient diffusion up to macroscopic viscosity. Based on a comparative discussion with results from other crystallin proteins, we suggest an interesting common pathway for dynamical arrest in all crystallin proteins, with potential implications for the understanding of crowding effects in the eye lens.

Neutron scattering elucidates lipid membrane dynamics

Neutron spin echo spectroscopy allows researchers to better understand the way soft matter like lipids moves during temperature changes.

Experimental test of the rotating magnetic field neutron spin-echo technique: Spin-echo and MIEZE for large energy transfers

Here we are describing the experimental realization of the rotating magnetic field neutron spin echo (RFSE) technique that is based on spin turners using truly rotating magnetic fields. RFSE has no practical limit for short Fourier times and a “shorty” instrument mode built on this principle can be easily used at generic neutron spin echo spectrometers to increase their dynamical range beyond that provided by currently used methods.The upper limit for Fourier times is estimated. The tripling of the field integral (and hence the maximum Fourier time) by means of the bootstrap method is experimentally demonstrated. The suitability of RFSE for the use of Fresnel-type correction elements aiming to equalize the field integral for divergent neutron beams scattered by the sample is shown. The experimental realization of the MIEZE method using the RFSE technique allowing for studies of depolarizing samples and/or with depolarizing sample environments is presented.

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.

Observation of TOF–MIEZE Signals with Focusing Mirrors at BL06, MLF, J-PARC

A neutron spin echo experiment for tiny samples with high energy resolution can be realized by combining modulated intensity by zero effort (MIEZE) technique and ellipsoidal neutron focusing supermirrors. We observed TOF–MIEZE (the time-of-flight method with MIEZE) signals with an effective frequency of 108 kHz with focusing beam. The Fourier time range 0.48–5.3 ns was covered at once by using TOF technique with neutron wavelength 0.65–1.45 nm. The contrasts of TOF–MIEZE signal were 0.62 ± 0.02 and 0.50 ± 0.05 with each neutron wavelength of 0.68 nm and 1.35 nm, respectively.

Structure and Dynamics of Irreversible Single-Chain Nanoparticles in Dilute Solution. A Neutron Scattering Investigation

We present a small-angle neutron scattering (SANS) and neutron spin echo investigation on the structure and dynamics of irreversible single-chain nanoparticles (SCNPs) in dilute solution. SCNPs are...

Enhanced dynamics in the anomalous melting regime of DMPG lipid membranes.

Like many soft materials, lipids undergo a melting transition associated with a significant increase in their dynamics. At temperatures below the main melting transition (Tm), all molecular and collective dynamics are suppressed, while above Tm the alkyl tail motions, lipid diffusivity, and collective membrane undulations are at least an order of magnitude faster. Here we study the collective dynamics of dimyristoylphosphatidylglycerol (DMPG, di 14:0 PG) using neutron spin echo spectroscopy throughout its anomalous phase transition that occurs over a 12 °C–20° C wide temperature window. Our results reveal that the membranes are softer and more dynamic during the phase transition than at higher temperatures corresponding to the fluid phase and provide direct experimental evidence for the predicted increase in membrane fluctuations during lipid melting. These results provide new insights into the nanoscale lipid membrane dynamics during the melting transition and demonstrate how these dynamics are coupled to changes in the membrane structure.