“Molecular Dynamics Simulation of an Engineered T4 Lysozyme Protein with Potential Nano-Biotechnological Applications”

The idea of developing a Bio-molecular mechanical device has attracted the attention of many research groups due to its promising Nano and Bio- technological applications such as flow-control valves, switches and bio-sensors. T4 Lysozyme protein is an enzyme, which has been extensively studied. This protein provides a wealth of information regarding the structure/function relationship of proteins at atomic resolution. An engineered variant of T4 Lysozyme has been reported to trigger a large scale translocation of an engineered helix (∼ 2nm), upon the addition of an external ligand. The design was based on the duplication of a surface helix, followed by manipulating the stability of an adjacent loop, which caused the duplicated helix to switch between two conformations. The purpose of this study is to investigate the dynamics of the engineered motion for potential Nano and Biotechnological applications. Many wild type and mutant static crystal structures of T4 Lysozyme have been shown to display a range of about 50 degrees in hinge bending motion between the N- and C-terminal domains of the protein, indicating intrinsic flexibility. The present molecular dynamics simulations detect similar motion in the engineered protein, within 100 nanosecond time scale. A preliminary mathematical model (solvent free) describing the hinge bending motion and the impact force is constructed. However, in the nanosecond time scale, the engineered triggered motion (the helical trans location) was partially detected only when bond constraints were significantly relaxed

The denatured state of N-PGK is compact and predominantly disordered

The Organisation of the structure present in the chemically denatured N-terminal domain of phosphoglycerate kinase (N-PGK) has been determined by paramagnetic relaxation enhancements (PREs) to define the conformational landscape accessible to the domain. Below 2.0 M guanidine hydrochloride (GuHCl), a species of N-PGK (denoted I-b) is detected, distinct from those previously characterised by kinetic experiments [folded (F), kinetic intermediate (I-k) and denatured (D)]. The transition to I-b is never completed at equilibrium, because F predominates below 1.0 M GuHCl. Therefore, the ability of PREs to report on transient or low population species has been exploited to characterise I-b. Five single cysteine variants of N-PGK were labelled with the nitroxide electron spin-label MTSL [(1-oxyl-2,2,5,5-tetramethyl-3-pyrroline-3-methyl)methanesulfonate] and the denaturant dependences of the relaxation properties of the amide NMR signals between 1.2 and 3.6 M GuHCl were determined. Significant PREs for I-b were obtained, but these were distributed almost uniformly throughout the sequence. Furthermore, the PREs indicate that no specific short tertiary contacts persist. The data indicate a collapsed state with no coherent three-dimensional structure, but with a restricted radius beyond which the protein chain rarely reaches. The NMR characteristics Of I-b indicate that it forms from the fully denatured state within 100 mu s, and therefore a rapid collapse is the initial stage of folding of N-PGK from its chemically denatured state. By extrapolation, I-b is the predominant form of the denatured state under native conditions, and the non-specifically collapsed structure implies that many non-native contacts and chain reversals form early in protein folding and must be broken prior to attaining the native state topology. (C) 2008 Elsevier Ltd. All rights reserved

Polarisable force fields: what do they add in biomolecular simulations?

The quality of biomolecular simulations critically depends on the accuracy of the force field used to calculate the potential energy of the molecular configurations. Currently, most simulations employ non-polarisable force fields, which describe electrostatic interactions as the sum of Coulombic interactions between fixed atomic charges. Polarisation of these charge distributions is incorporated only in a mean-field manner. In the past decade, extensive efforts have been devoted to developing simple, efficient, and yet generally applicable polarisable force fields for biomolecular simulations. In this review, we summarise the latest developments in accounting for key biomolecular interactions with polarisable force fields and applications to address challenging biological questions. In the end, we provide an outlook for future development in polarisable force fields

Polarisable force fields:what do they add in biomolecular simulations?

The quality of biomolecular simulations critically depends on the accuracy of the force field used to calculate the potential energy of the molecular configurations. Currently, most simulations employ non-polarisable force fields, which describe electrostatic interactions as the sum of Coulombic interactions between fixed atomic charges. Polarization of these charge distributions is incorporated only in a mean-field manner. In the past decade, extensive efforts have been devoted to developing simple, efficient, and yet generally applicable polarisable force fields for biomolecular simulations. In this review, we summarise the latest developments in accounting for key biomolecular interactions with polarisable force fields and applications to address challenging biological questions. In the end, we provide an outlook for future development in polarisable force fields.Comment: 25 pages, 3 figure

Protein–Mineral Interactions: Molecular Dynamics Simulations Capture Importance of Variations in Mineral Surface Composition and Structure

Molecular dynamics simulations, conventional and metadynamics, were performed to determine the interaction of model protein Gb1 over kaolinite (001), Na⁺-montmorillonite (001), Ca²⁺-montmorillonite (001), goethite (100), and Na⁺-birnessite (001) mineral surfaces. Gb1, a small (56 residue) protein with a well-characterized solution-state nuclear magnetic resonance (NMR) structure and having α-helix, 4-fold β-sheet, and hydrophobic core features, is used as a model protein to study protein soil mineral interactions and gain insights on structural changes and potential degradation of protein. From our simulations, we observe little change to the hydrated Gb1 structure over the kaolinite, montmorillonite, and goethite surfaces relative to its solvated structure without these mineral surfaces present. Over the Na⁺ -birnessite basal surface, however, the Gb1 structure is highly disturbed as a result of interaction with this birnessite surface. Unraveling of the Gb1 β-sheet at specific turns and a partial unraveling of the α-helix is observed over birnessite, which suggests specific vulnerable residue sites for oxidation or hydrolysis possibly leading to fragmentation

Computational Studies of Molecular Mechanisms Mediating Protein Adsorption on Material Surfaces

Protein adsorption at material surfaces is a fundamental concept in many scientific applications ranging from the biocompatibility of implant materials in bioengineering to cleaning environmental material surfaces from toxic proteins in the area of biodefense. Understanding the molecular-level details of protein-surface interactions is crucial for controlling protein adsorption. While a range of experimental techniques has been developed to study protein adsorption, these techniques cannot produce the fundamental molecular-level information of protein adsorption. All-atom empirical force field molecular dynamics (MD) simulations hold great promise as a valuable tool for elucidating and predicting the mechanisms governing protein adsorption. However, current MD simulation methods have not been validated for this application. This research addresses three limitations of the standard MD when applied to the simulations of the protein-surface interactions: (1) representation of the force field parameters governing the interactions of protein amino acids with the material surface; (2) cluster analysis of ensembles of adsorbed protein states obtained in protein-adsorption simulations, in which in addition to the conformation the orientation of the sampled states is also important; and (3) simulation time to ensure a significant level of conformational sampling to cover the entire rough energy landscape of such a large molecular system as protein adsorption. This study, thus, attempted to further advance protein-adsorption simulation methods using high-density polyethylene as a model materials surface

I. Dissociation free energies of drug-receptor systems: Via non-equilibrium alchemical simulations: A theoretical framework

In this contribution I critically revise the alchemical reversible approach in the context of the statistical mechanics theory of non covalent bonding in drug receptor systems. I show that most of the pitfalls and entanglements for the binding free energies evaluation in computer simulations are rooted in the equilibrium assumption that is implicit in the reversible method. These critical issues can be resolved by using a non-equilibrium variant of the alchemical method in molecular dynamics simulations, relying on the production of many independent trajectories with a continuous dynamical evolution of an externally driven alchemical coordinate, completing the decoupling of the ligand in a matter of few tens of picoseconds rather than nanoseconds. The absolute binding free energy can be recovered from the annihilation work distributions by applying an unbiased unidirectional free energy estimate, on the assumption that any observed work distribution is given by a mixture of normal distributions, whose components are identical in either direction of the non-equilibrium process, with weights regulated by the Crooks theorem. I finally show that the inherent reliability and accuracy of the unidirectional estimate of the decoupling free energies, based on the production of few hundreds of non-equilibrium independent sub-nanoseconds unrestrained alchemical annihilation processes, is a direct consequence of the funnel-like shape of the free energy surface in molecular recognition. An application of the technique on a real drug-receptor system is presented in the companion paper.Comment: 34 pages, 4 figure

並列カスケード選択分子動力学法による生体分子の会合・解離シミュレーション

学位の種別: 課程博士審査委員会委員 : （主査）東京大学教授 津田 宏治, 東京大学客員准教授 広川 貴次, 東京大学特任准教授 寺田 透, 東京大学講師 笠原 雅弘, 東京大学准教授 木立 尚孝University of Tokyo(東京大学

Development of High Performance Molecular Dynamics with Application to Multimillion-Atom Biomass Simulations

An understanding of the recalcitrance of plant biomass is important for efficient economic production of biofuel. Lignins are hydrophobic, branched polymers and form a residual barrier to effective hydrolysis of lignocellulosic biomass. Understanding lignin\u27s structure, dynamics and its interaction and binding to cellulose will help with finding more efficient ways to reduce its contribution to the recalcitrance. Molecular dynamics (MD) using the GROMACS software is employed to study these properties in atomic detail. Studying complex, realistic models of pretreated plant cell walls, requires simulations significantly larger than was possible before. The most challenging part of such large simulations is the computation of the electrostatic interaction. As a solution, the reaction-field (RF) method has been shown to give accurate results for lignocellulose systems, as well as good computational efficiency on leadership class supercomputers. The particle-mesh Ewald method has been improved by implementing 2D decomposition and thread level parallelization for molecules not accurately modeled by RF. Other scaling limiting computational components, such as the load balancing and memory requirements, were identified and addressed to allow such large scale simulations for the first time. This work was done with the help of modern software engineering principles, including code-review, continuous integration, and integrated development environments. These methods were adapted to the special requirements for scientific codes. Multiple simulations of lignocellulose were performed. The simulation presented primarily, explains the temperature-dependent structure and dynamics of individual softwood lignin polymers in aqueous solution. With decreasing temperature, the lignins are found to transition from mobile, extended to glassy, compact states. The low-temperature collapse is thermodynamically driven by the increase of the translational entropy and density fluctuations of water molecules removed from the hydration shell