Minimum energy paths for conformational changes of viral capsids

In this work we study how a viral capsid can change conformation using techniques of Large Deviations Theory for stochastic differential equations. The viral capsid is a model of a complex system in which many units - the proteins forming the capsomers - interact by weak forces to form a structure with exceptional mechanical resistance. The destabilization of such a structure is interesting both per se, since it is related either to infection or maturation processes, and because it yields insights into the stability of complex structures in which the constitutive elements interact by weak attractive forces. We focus here on a simplified model of a dodecahederal viral capsid, and assume that the capsomers are rigid plaquettes with one degree of freedom each. We compute the most probable transition path from the closed capsid to the final configuration using minimum energy paths, and discuss the stability of intermediate states.Comment: 27 pages, 4 figures. New version, to appear in Physical Review

Protein container disassembly pathways depend on geometric design

The majority of viruses are organised according to the structural blueprints of the seminal Caspar-Klug theory. However, there are a number of notable exceptions to this geometric design principle. Prominent examples are the cancer-causing papilloma viridae and the \textit{de novo} designed AaLS cages that exhibit non-quasiequivalent capsid structures with protein numbers excluded by Caspar-Klug theory. The biophysical properties of these geometrically distinct architectures and the fitness advantages driving their evolution are currently unclear. We investigate here the resilience to fragmentation and disassembly behaviour of these capsid geometries by introducing a percolation theory on weighted graphs. We show that these cage architectures follow one of two distinct disassembly pathways, preferring either hole formation or capsid fragmentation. This suggests that preference for specific disassembly scenarios could be a driving force for the evolution of the non Caspar-Klug protein container architectures.Comment: 15 pages, 10 figure

Bioengineering Strategies for Protein-Based Nanoparticles.

In recent years, the practical application of protein-based nanoparticles (PNPs) has expanded rapidly into areas like drug delivery, vaccine development, and biocatalysis. PNPs possess unique features that make them attractive as potential platforms for a variety of nanobiotechnological applications. They self-assemble from multiple protein subunits into hollow monodisperse structures; they are highly stable, biocompatible, and biodegradable; and their external components and encapsulation properties can be readily manipulated by chemical or genetic strategies. Moreover, their complex and perfect symmetry have motivated researchers to mimic their properties in order to create de novo protein assemblies. This review focuses on recent advances in the bioengineering and bioconjugation of PNPs and the implementation of synthetic biology concepts to exploit and enhance PNP's intrinsic properties and to impart them with novel functionalities

Hybrid organic-inorganic nanoparticles: controlled incorporation of gold nanoparticles into virus-like particles and application in surface-enhanced Raman spectroscopy.

A capsid is the protein coat surrounding a virus' genome that ensures its protection and transport. The capsid of murine polyomavirus (muPy) consists of one major (VP1) and two minor (VP2/3) proteins, from which just VP1 is sufficient to form the capsid when expressed recombinantly (1). From a material engineering point of view, viral capsids are of interest because they present a paradigm for complex self-assembly on the nanometer scale. Understanding and controlling these assembly dynamics will allow the construction of nanoscale structures using a self-assembly process. The first step in this direction was the discovery that capsids of several viruses can be reversibly disassembled into their building blocks and reassembled using the same building blocks by simply changing the buffer conditions (2, 3). Such capsids already find applications as targeted in vivo delivery vectors for genes, proteins or small molecular drugs (4, 5), as optical probes for biomedical imaging and sensing purposes with unprecedented resolution and sensitivity and can potentially be used as templates for nanoelectronics (6, 7)

Biophysical properties of single rotavirus particles account for the functions of protein shells in a multilayered virus

The functions performed by the concentric shells of multilayered dsRNA viruses require specific protein interactions that can be directly explored through their mechanical properties. We studied the stiffness, breaking force, critical strain and mechanical fatigue of individual Triple, Double and Single layered rotavirus (RV) particles. Our results, in combination with Finite Element simulations, demonstrate that the mechanics of the external layer provides the resistance needed to counteract the stringent conditions of extracellular media. Our experiments, in combination with electrostatic analyses, reveal a strong interaction between the two outer layers and how it is suppressed by the removal of calcium ions, a key step for transcription initiation. The intermediate layer presents weak hydrophobic interactions with the inner layer that allow the assembly and favor the conformational dynamics needed for transcription. Our work shows how the biophysical properties of the three shells are finely tuned to produce an infective RV virio

On the Effect of Thermodynamic Equilibrium on the Assembly Efficiency of Complex Multi-Layered Virus-Like Particles (VLP): the Case of Rotavirus VLP

Previous studies have reported the production of malformed virus-like-particles (VLP) in recombinant host systems. Here we computationally investigate the case of a large triple-layered rotavirus VLP (RLP). In vitro assembly, disassembly and reassembly data provides strong evidence of microscopic reversibility of RLP assembly. Light scattering experimental data also evidences a slow and reversible assembly untypical of kinetic traps, thus further strengthening the fidelity of a thermodynamically controlled assembly. In silico analysis further reveals that under favourable conditions particles distribution is dominated by structural subunits and completely built icosahedra, while other intermediates are present only at residual concentrations. Except for harshly unfavourable conditions, assembly yield is maximised when proteins are provided in the same VLP protein mass composition. The assembly yield decreases abruptly due to thermodynamic equilibrium when the VLP protein mass composition is not obeyed. The latter effect is more pronounced the higher the Gibbs free energy of subunit association is and the more complex the particle is. Overall this study shows that the correct formation of complex multi-layered VLPs is restricted to a narrow range of association energies and protein concentrations, thus the choice of the host system is critical for successful assembly. Likewise, the dynamic control of intracellular protein expression rates becomes very important to minimize wasted proteins