Tilo Seydel

Protein self-diffusion in crowded solutions

Macromolecular crowding in biological media is an essential factor for cellular function. The interplay of intermolecular interactions at multiple time and length scales governs a fine-tuned system of reaction and transport processes, including particularly protein diffusion as a limiting or driving factor. Using quasielastic neutron backscattering, we probe the protein self-diffusion in crowded aqueous solutions of bovine serum albumin on nanosecond time and nanometer length scales employing the same protein as crowding agent. The measured diffusion coefficient D(φ) strongly decreases with increasing protein volume fraction φ explored within 7% ≤ φ ≤ 30%. With an ellipsoidal protein model and an analytical framework involving colloid diffusion theory, we separate the rotational Dr(φ) and translational Dt(φ) contributions to D(φ). The resulting Dt(φ) is described by short-time self-diffusion of effective spheres. Protein self-diffusion at biological volume fractions is found to be slowed down to 20% of the dilute limit solely due to hydrodynamic interactions.

Molecular motions in lipid bilayers studied by the neutron backscattering technique.

We report a high energy-resolution neutron backscattering study to investigate slow motions on nanosecond time scales in highly oriented solid supported phospholipid bilayers of the model system DMPC-d54 (deuterated 1,2-dimyristoyl-sn-glycero-3-phoshatidylcholine), hydrated with heavy water. This technique allows to discriminate the onset of mobility at different length scales for the different molecular components, as, e.g., the lipid acyl-chains and the hydration water in between the membrane stacks, respectively, and provides a benchmark test regarding the feasibility of neutron backscattering investigations on these sample systems. We discuss freezing of the lipid acyl-chains, as observed by this technique, and observe a second freezing transition which we attribute to the hydration water.

Viscosity and diffusion: crowding and salt effects in protein solutions

We report on a joint experimental–theoretical study of collective diffusion in, and static shear viscosity of solutions of bovine serum albumin (BSA) proteins, focusing on the dependence on protein and salt concentration. Data obtained from dynamic light scattering and rheometric measurements are compared to theoretical calculations based on an analytically treatable spheroid model of BSA with isotropic screened Coulomb plus hard-sphere interactions. The only input to the dynamics calculations is the static structure factor obtained from a consistent theoretical fit to a concentration series of small-angle X-ray scattering (SAXS) data. This fit is based on an integral equation scheme that combines high accuracy with low computational cost. All experimentally probed dynamic and static properties are reproduced theoretically with an at least semi-quantitative accuracy. For lower protein concentration and low salinity, both theory and experiment show a maximum in the reduced viscosity, caused by the electrostatic repulsion of proteins. On employing our theoretical and experimental results, the applicability range of a generalized Stokes–Einstein (GSE) relation connecting viscosity, collective diffusion coefficient, and osmotic compressibility, proposed by Kholodenko and Douglas [Phys. Rev. E, 1995, 51, 1081] is examined. Significant violation of the GSE relation is found, both in experimental data and in theoretical models, in concentrated systems at physiological salinity, and under low-salt conditions for arbitrary protein concentrations.

Motional Coherence in Fluid Phospholipid Membranes

We report a high energy-resolution neutron backscattering study, combined with in situ diffraction, to investigate slow molecular motions on nanosecond time scales in the fluid phase of phospholipid bilayers of 1,2-dimyristoyl-sn-glycero-3-phoshatidylcholine. A cooperative structural relaxation process was observed. From the in-plane scattering vector dependence of the relaxation rates in hydrogenated and deuterated samples, combined with results from a 0.1 micros long all-atom molecular dynamics simulation, it is concluded that correlated dynamics in lipid membranes occurs over several lipid distances, spanning a time interval from pico- to nanoseconds.

Protein diffusion in crowded electrolyte solutions.

We report on a combined cold neutron backscattering and spin-echo study of the short-range and long-range nanosecond diffusion of the model globular protein bovine serum albumin (BSA) in aqueous solution as a function of protein concentration and NaCl salt concentration. Complementary small angle X-ray scattering data are used to obtain information on the correlations of the proteins in solution. Particular emphasis is put on the effect of crowding, i.e. conditions under which the proteins cannot be considered as objects independent of each other. We thus address the question at which concentration this crowding starts to influence the static and in particular also the dynamical behaviour. We also briefly discuss qualitatively which charge effects, i.e. effects due to the interplay of charged molecules in an electrolyte solution, may be anticipated. Both the issue of crowding as well as that of charge effects are particularly relevant for proteins and their function under physiological conditions, where the protein volume fraction can be up to approximately 40% and salt ions are ubiquitous. The interpretation of the data is put in the context of existing studies on related systems and of existing theoretical models.

Nanosecond molecular relaxations in lipid bilayers studied by high energy-resolution neutron scattering and in situ diffraction

We report a high energy-resolution neutron backscattering study to investigate slow motions on nanosecond time scales in highly oriented solid-supported phospholipid bilayers of the model system deuterated 1,2-dimyristoyl-sn-glycero-3-phosphatidylcholine, hydrated with heavy water. Wave-vector-resolved quasielastic neutron scattering is used to determine relaxation times tau , which can be associated with different molecular components, i.e., the lipid acyl chains and the interstitial water molecules in the different phases of the model membrane system. The inelastic data are complemented by both energy-resolved and energy-integrated in situ diffraction. From a combined analysis of the inelastic data in the energy and time domains, the corresponding character of the relaxation, i.e., the exponent of the exponential decay, is also determined. From this analysis we quantify two relaxation processes. We associate the fast relaxation with translational diffusion of lipid and water molecules while the slow process likely stems from collective dynamics.

Diffusion and dynamics of γ-globulin in crowded aqueous solutions.

Dynamics in protein solutions is essential for both protein function and cellular processes. The hierarchical complexity of global protein diffusion, side-chain diffusion, and microscopic motions of chemical groups renders a complete understanding challenging. We present results from quasi-elastic neutron scattering on protein solutions of γ-globulin over a wide range of volume fractions. Translational and rotational diffusion can be self-consistently separated from internal motions. The global diffusion is consistent with predictions for effective spheres even though the branched molecular shape differs considerably from a colloidal sphere. The internal motions are characterized both geometrically and dynamically, suggesting a picture of methyl rotations and restricted diffusion of side chains. We show that the advent of new neutron spectrometers allows the study of current questions including the coupling of intracellular dynamics and protein function.

Short range ballistic motion in fluid lipid bilayers studied by quasi-elastic neutron scattering

Diffusion is the primary mechanism for movement of lipids and proteins in the lateral direction of a biological membrane. In this paper we have used quasi-elastic neutron scattering to examine the diffusion process of lipid molecules in fluid DMPC membranes. We found that the motion over length scales greater than the lipid diameter could be characterized as a continuous diffusion process, with a diffusion coefficient of D = 64 × 10−12 m2/s. The continuous diffusion model has been successfully used in the past to describe the motion of lipid over long length scales. However, the focus of this measurement was to determine how the character of the molecular motion changes on length scales shorter than the nearest neighbour distance. At very short length scales (<2.37 A), we see first experimental evidence for a short-range flow-like ballistic motion.

Exploring the collective dynamics of lipid membranes with inelastic neutron scattering

While most spectroscopic techniques, as e.g., nuclear magnetic resonance or dielectric spectroscopy, probe macroscopic responses, neutron and within some restrictions also x-ray scattering experiments give the unique access to microscopic dynamics at length scales of intermolecular or atomic distances. Only recently, it has become possible to study collective dynamics of planar lipid bilayers using neutron spectroscopy techniques [M. Rheinstadter, C. Ollinger, G. Fragneto, F. Demmel, and T. Salditt, Phys. Rev. Lett. 93, 108107 (2004)]. We determined the dispersion relation of the coherent fast picosecond density fluctuations on nearest-neighbor distances of the phospholipid acyl chains in the gel and in the fluid phases of a dimyristoylphoshatidylcholine bilayer. The experiments shed light on the evolution of structure and dynamics, and the relation between them, in the range of the gel-fluid main phase transition. The scattering volume restriction for inelastic neutron experiments was overcome by stackin...

How mobile are protons in the structure of dental glass ionomer cements

The development of dental materials with improved properties and increased longevity can save costs and minimize discomfort for patients. Due to their good biocompatibility, glass ionomer cements are an interesting restorative option. However, these cements have limited mechanical strength to survive in the challenging oral environment. Therefore, a better understanding of the structure and hydration process of these cements can bring the necessary understanding to further developments. Neutrons and X-rays have been used to investigate the highly complex pore structure, as well as to assess the hydrogen mobility within these cements. Our findings suggest that the lower mechanical strength in glass ionomer cements results not only from the presence of pores, but also from the increased hydrogen mobility within the material. The relationship between microstructure, hydrogen mobility and strength brings insights into the materials durability, also demonstrating the need and opening the possibility for further research in these dental cements.