Quantitative approaches for characterising fibrillar protein nanostructures

Polypeptide sequences have an inherent tendency to self-assemble into filamentous nanostructures commonly known as amyloid fibrils. Such self-assembly is used in nature to generate a variety of functional materials ranging from protective coatings in bacteria to catalytic scaffolds in mammals. The aberrant self-assembly of misfolded peptides and proteins is also, however, implicated in a range of disease states including neurodegenerative conditions such as Alzheimer's and Parkinson's diseases. It is increasingly evident that the intrinsic material properties of these structures are crucial for understanding the thermodynamics and kinetics of the pathological deposition of proteins, particularly as the mechanical fragmentation of aggregates enhances the rate of protein deposition by exposing new fibril ends which can promote further growth. We discuss here recent advances in physical techniques that are able to characterise the hierarchical self-assembly of misfolded protein molecnles and define their properties. © 2010 Materials Research Society.

Perturbation of the stability of amyloid fibrils through alteration of electrostatic interactions.

The self-assembly of proteins and peptides into polymeric amyloid fibrils is a process that has important implications ranging from the understanding of protein misfolding disorders to the discovery of novel nanobiomaterials. In this study, we probe the stability of fibrils prepared at pH 2.0 and composed of the protein insulin by manipulating electrostatic interactions within the fibril architecture. We demonstrate that strong electrostatic repulsion is sufficient to disrupt the hydrogen-bonded, cross-β network that links insulin molecules and ultimately results in fibril dissociation. The extent of this dissociation correlates well with predictions for colloidal models considering the net global charge of the polypeptide chain, although the kinetics of the process is regulated by the charge state of a single amino acid. We found the fibrils to be maximally stable under their formation conditions. Partial disruption of the cross-β network under conditions where the fibrils remain intact leads to a reduction in their stability. Together, these results support the contention that a major determinant of amyloid stability stems from the interactions in the structured core, and show how the control of electrostatic interactions can be used to characterize the factors that modulate fibril stability.

Far-infrared emission and absorption by hot carriers in superlattices

Some of the earliest theoretical speculation, stimulated by the growth of semiconductor superlattices, focused on novel devices based on vertical transport through engineered band structures; Esaki and Tsu promised Bloch oscillators in narrow mini-band systems and Kazarinov and Suris contemplated electrically stimulated intersubband transitions as sources of infrared radiation. Nearly twenty years later these material systems have been perfected, characterized and understood and experiments are emerging that test some of these original concepts for novel submillimetre wave electronics. Here we describe recent experiments on intersubband emission in quantum wells stimulated by resonant tunnelling currents. A critical issue at this time is devising a way to achieve population inversion. Other experiments explore 'saturation' effects in narrow miniband transport. Thermal saturation may be viewed as a precursor to Bloch oscillation if the same effects can be induced with an applied electric field.

Twisting transition between crystalline and fibrillar phases of aggregated peptides

We study two distinctly ordered condensed phases of polypeptide molecules, amyloid fibrils and amyloidlike microcrystals, and the first-order twisting phase transition between these two states. We derive a single free-energy form which connects both phases. Our model identifies relevant degrees of freedom for describing the collective behavior of supramolecular polypeptide structures, reproduces accurately the results from molecular dynamics simulations as well as from experiments, and sheds light on the uniform nature of the dimensions of different peptide fibrils. © 2012 American Physical Society.

Electrostatic effects in filamentous protein aggregation.

Electrostatic forces play a key role in mediating interactions between proteins. However, gaining quantitative insights into the complex effects of electrostatics on protein behavior has proved challenging, due to the wide palette of scenarios through which both cations and anions can interact with polypeptide molecules in a specific manner or can result in screening in solution. In this article, we have used a variety of biophysical methods to probe the steady-state kinetics of fibrillar protein self-assembly in a highly quantitative manner to detect how it is modulated by changes in solution ionic strength. Due to the exponential modulation of the reaction rate by electrostatic forces, this reaction represents an exquisitely sensitive probe of these effects in protein-protein interactions. Our approach, which involves a combination of experimental kinetic measurements and theoretical analysis, reveals a hierarchy of electrostatic effects that control protein aggregation. Furthermore, our results provide a highly sensitive method for the estimation of the magnitude of binding of a variety of ions to protein molecules.

Magnetic shielding properties of high- Tc superconducting hollow cylinders: Model combining experimental data for axial and transverse magnetic field configurations

Magnetic shielding efficiency was measured on high- Tc superconducting hollow cylinders subjected to either an axial or a transverse magnetic field in a large range of field sweep rates, dBapp/dt. The behaviour of the superconductor was modelled in order to reproduce the main features of the field penetration curves by using a minimum number of free parameters suitable for both magnetic field orientations. The field penetration measurements were carried out on Pb-doped Bi-2223 tubes at 77K by applying linearly increasing magnetic fields with a constant sweep rate ranging between 10νTs-1 and 10mTs-1 for both directions of the applied magnetic field. The experimental curves of the internal field versus the applied field, Bin(Bapp), show that, at a given sweep rate, the magnetic field for which the penetration occurs, Blim, is lower for the transverse configuration than for the axial configuration. A power law dependence with large exponent, n′, is found between Blim and dBapp/dt. The values of n′ are nearly the same for both configurations. We show that the main features of the curves B in(Bapp) can be reproduced using a simple 2D model, based on the method of Brandt, involving a E(J) power law with an n-exponent and a field-dependent critical current density, Jc(B), (following the Kim model: Jc = Jc0(1+B/B1)-1). In particular, a linear relationship between the measured n′-exponents and the n-exponent of the E(J) power law is suggested by taking into account the field dependence of the critical current density. Differences between the axial and the transverse shielding properties can be simply attributed to demagnetizing fields. © 2009 IOP Publishing Ltd.

Transient ventilation dynamics induced by heat sources of unequal strength

We examine theoretically the transient displacement flow and density stratification that develops within a ventilated box after two localized floor-level heat sources of unequal strengths are activated. The heat input is represented by two non-interacting turbulent axisymmetric plumes of constant buoyancy fluxes B1 and B2 > B1. The box connects to an unbounded quiescent external environment of uniform density via openings at the top and base. A theoretical model is developed to predict the time evolution of the dimensionless depths λj and mean buoyancies δj of the 'intermediate' (j = 1) and 'top' (j = 2) layers leading to steady state. The flow behaviour is classified in terms of a stratification parameter S, a dimensionless measure of the relative forcing strengths of the two buoyant layers that drive the flow. We find that dδ1/dτ α 1/λ1 and dδ2/dτ α 1/λ2, where τ is a dimensionless time. When S 1, the intermediate layer is shallow (small λ1), whereas the top layer is relatively deep (large λ2) and, in this limit, δ1 and δ2 evolve on two characteristically different time scales. This produces a time lag and gives rise to a 'thermal overshoot', during which δ1 exceeds its steady value and attains a maximum during the transients; a flow feature we refer to, in the context of a ventilated room, as 'localized overheating'. For a given source strength ratio ψ = B1/B2, we show that thermal overshoots are realized for dimensionless opening areas A < Aoh and are strongly dependent on the time history of the flow. We establish the region of {A, ψ} space where rapid development of δ1 results in δ1 > δ2, giving rise to a bulk overturning of the buoyant layers. Finally, some implications of these results, specifically to the ventilation of a room, are discussed. © Cambridge University Press 2013.