Molecular Modeling on Inhibitor Complexes and Active-Site Dynamics of Cytochrome P450 C17, a Target for Prostate Cancer Therapy

A molecular model for the P450 enzyme cytochrome P450 C17 (CYP17) is presented based on sequence alignments of multiple template structures and homology modeling. This enzyme plays a central role in the biosynthesis of testosterone and is emerging as a major target in prostate cancer, with the recently developed inhibitor abiraterone currently in advanced clinical trials. The model is described in detail, together with its validation, by providing structural explanations to available site-directed mutagenesis data. The CYP17 molecule in this model is in the form of a triangular prism, with an edge of similar to 55 angstrom and a thickness of similar to 37 angstrom. It is predominantly helical, comprising 13 alpha helices interspersed by six 3(10) helices and 11 beta-sheets. Multinanosecond molecular dynamics simulations in explicit solvent have been carried out, and principal components analysis has been used to reveal the details of dynamics around the active site. Coarse-grained methods have also been used to verify low-frequency motions, which have been correlated with active-site gating. The work also describes the results of docking synthetic inhibitors, including the drug abiraterone and the natural substrate pregnenolone, in the CYP17 active site together with molecular dynamics simulations on the complexes. (C) 2010 Elsevier Ltd. All rights reserved.

Molecular dynamics and principal components analysis of human telomeric quadruplex multimers.

Guanine-rich DNA repeat sequences located at the terminal ends of chromosomal DNA can fold in a sequence-dependent manner into G-quadruplex structures, notably the terminal 150–200 nucleotides at the 3' end, which occur as a single-stranded DNA overhang. The crystal structures of quadruplexes with two and four human telomeric repeats show an all-parallel-stranded topology that is readily capable of forming extended stacks of such quadruplex structures, with external TTA loops positioned to potentially interact with other macromolecules. This study reports on possible arrangements for these quadruplex dimers and tetramers, which can be formed from 8 or 16 telomeric DNA repeats, and on a methodology for modeling their interactions with small molecules. A series of computational methods including molecular dynamics, free energy calculations, and principal components analysis have been used to characterize the properties of these higher-order G-quadruplex dimers and tetramers with parallel-stranded topology. The results confirm the stability of the central G-tetrads, the individual quadruplexes, and the resulting multimers. Principal components analysis has been carried out to highlight the dominant motions in these G-quadruplex dimer and multimer structures. The TTA loop is the most flexible part of the model and the overall multimer quadruplex becoming more stable with the addition of further G-tetrads. The addition of a ligand to the model confirms the hypothesis that flat planar chromophores stabilize G-quadruplex structures by making them less flexible.

Framework for Establishing Limits of Tabular Aerodynamic Models for Flight Dynamics Analysis

This paper describes the use of the Euler equations for the generation and testing of tabular aerodynamic models for flight dynamics analysis. Maneuvers for the AGARD Standard Dynamics Model sharp leading-edge wind-tunnel geometry are considered as a test case. Wind-tunnel data is first used to validate the prediction of static and dynamic coefficients at both low and high angles, featuring complex vortical flow, with good agreement obtained at low to moderate angles of attack. Then the generation of aerodynamic tables is described based on a data fusion approach. Time-optimal maneuvers are generated based on these tables, including level flight trim, pull-ups at constant and varying incidence, and level and 90 degrees turns. The maneuver definition includes the aircraft states and also the control deflections to achieve the motion. The main point of the paper is then to assess the validity of the aerodynamic tables which were used to define the maneuvers. This is done by replaying them, including the control surface motions, through the time accurate computational fluid dynamics code. The resulting forces and moments are compared with the tabular values to assess the presence of inadequately modeled dynamic or unsteady effects. The agreement between the tables and the replay is demonstrated for slow maneuvers. Increasing rate maneuvers show discrepancies which are ascribed to vortical flow hysteresis at the higher rate motions. The framework is suitable for application to more complex viscous flow models, and is powerful for the assessment of the validity of aerodynamics models of the type currently used for studies of flight dynamics.

Local Conentration of Waves due to Abrupt Alongshore Variation of Nearshore Bathymetry

Small-scale physical and numerical experiments were conducted to investigate the local concentration of waves (monochromatic and group) due to abrupt change of nearshore bathymetry in alongshore direction. Wave run-up motions along the shoreline were measured using an image analysis technique to compare localized concentration of wave energy, when waves propagate a over bathymetry composing rhythmic patterns of mild/steep slope bottom configurations. Measured alongshore variation of maximum wave run-up heights showed significant peak near the boundary, which has sudden alongshore change of depth, both under monochromatic and group wave trains. This phenomenon is found to be due to interaction of waves with neashore currents, which is further enhanced by excitation of long wave components by breaking of group waves. Furthermore, this paper discusses results of preliminary experiments carried out to test the effectiveness of several shore protection structure layouts in mitigating such wave concentrations. Numerical simulations were performed by using a model developed based on Nwogu (1993) Boussinesq-type equations; coupled with a transport equation to model energy dissipation due to wave breaking.

Photospheric magnetic vortex structures

Using direct numerical magneto-hydrodynamic (MHD) simulations, we demonstrate the evidence of two physically different types of vortex motions in the solar photosphere. Baroclinic motions of plasma in non-magnetic granules are the primary source of vorticity in granular regions of the solar photosphere, however, there is a significantly more efficient mechanism of vorticity production in strongly magnetised intergranular lanes. These swirly motions of plasma in intergranular magnetic field concentrations could be responsible for the generation of different types of MHD wave modes, for example, kink, sausage and torsional Alfven waves. These waves could transport a relevant amount of energy from the lower solar atmosphere and contribute to coronal plasma heating.

Transverse Oscillations in Chromospheric Mottles

A number of recent investigations have revealed that transverse waves are ubiquitous in the solar chromosphere. The vast majority of these have been reported in limb spicules and active region fibrils. We investigate long-lived, quiet-Sun, on-disk features such as chromospheric mottles (jet-like features located at the boundaries of supergranular cells) and their transverse motions. The observations were obtained with the Rapid Oscillations in the Solar Atmosphere instrument at the Dunn Solar Telescope. The data set is comprised of simultaneous imaging in the Ha core, Ca II K, and G band of an on-disk quiet-Sun region. Time-distance techniques are used to study the characteristics of the transverse oscillations. We detect over 40 transverse oscillations in both bright and dark mottles, with periods ranging from 70 to 280 s, with the most frequent occurrence at ~165 s. The velocity amplitudes and transverse displacements exhibit characteristics similar to limb spicules. Neighboring mottles oscillating in-phase are also observed. The transverse oscillations of individual mottles are interpreted in terms of magnetohydrodynamic kink waves. Their estimated periods and damping times are consistent with phase mixing and resonant mode conversion.

Rapid Fluctuations in the Lower Solar Atmosphere

The Rapid Oscillations in the Solar Atmosphere instrument reveals solar atmospheric fluctuations at high frequencies. Spectra of variations of the G-band intensity (IG ) and Ca II K-line intensity (IK ) show correlated fluctuations above white noise to frequencies beyond 300 mHz and 50 mHz, respectively. The noise-corrected G-band spectrum for f = 28-326 mHz shows a power law with exponent -1.21 ± 0.02, consistent with the presence of turbulent motions. G-band spectral power in the 25-100 mHz ("UHF") range is concentrated at the locations of magnetic bright points in the intergranular lanes and is highly intermittent in time. The intermittence of the UHF G-band fluctuations, shown by a positive kurtosis ?, also suggests turbulence. Combining values of IG , IK , UHF power, and ? reveals two distinct states of the solar atmosphere. State 1, including almost all the data, is characterized by low IG , IK , and UHF power and ? ˜ 6. State 2, including only a very small fraction of the data, is characterized by high IG , IK , and UHF power and ? ˜ 3. Superposed epoch analysis shows that the UHF power peaks simultaneously with spatio-temporal IG maxima in either state. For State 1, IK shows 3.5 minute chromospheric oscillations with maxima occurring 21 s after IG maxima implying a 150-210 km effective height difference. However, for State 2 the IK and IG maxima are simultaneous; in this highly magnetized environment sites of G-band and K-line emission may be spatially close together.

Molecular Dynamics of HIV1-Integrase in Complex with 93del - A Structural Perspective on the Mechanism of Inhibition

HIV1 integrase is an important target for the antiviral therapy. Guanine-rich quadruplex, such as 93del, have been shown to be potent inhibitors of this enzyme and thus representing a new class of antiviral agents. Although X-ray and NMR structures of HIV1 integrase and 93del have been reported, there is no structural information of the complex and the mechanism of inhibition still remains unexplored. A number of computational methods including automated protein-DNA docking and molecular dynamics simulation in explicit solvent were used to model the binding of 93del to HIV1 integrase. Analysis of the dynamic behaviour of the complex using principal components analysis and elastic network modelling techniques allow us to understand how the binding of 93del aptamer and its interactions with key residues affect the intrinsic motions of the catalytic loops by stabilising them in catalytically inactive conformations. Such insights into the structural mechanism of inhibition can aid in improving the design of anti-HIV aptamers.

The molecular dynamics of Trypanosoma brucei UDP-galactose 4'-epimerase:a drug target for African sleeping sickness

During the past century, several epidemics of human African trypanosomiasis, a deadly disease caused by the protist Trypanosoma brucei, have afflicted sub-Saharan Africa. Over 10 000 new victims are reported each year, with hundreds of thousands more at risk. As current drug treatments are either highly toxic or ineffective, novel trypanocides are urgently needed. The T. brucei galactose synthesis pathway is one potential therapeutic target. Although galactose is essential for T. brucei survival, the parasite lacks the transporters required to intake galactose from the environment. UDP-galactose 4'-epimerase (TbGalE) is responsible for the epimerization of UDP-glucose to UDP-galactose and is therefore of great interest to medicinal chemists. Using molecular dynamics simulations, we investigate the atomistic motions of TbGalE in both the apo and holo states. The sampled conformations and protein dynamics depend not only on the presence of a UDP-sugar ligand, but also on the chirality of the UDP-sugar C4 atom. This dependence provides important insights into TbGalE function and may help guide future computer-aided drug discovery efforts targeting this protein.

Stellar Surface Magneto-Convection as a Source of Astrophysical Noise. I. Multi-component Parameterisation of Absorption Line Profiles

We outline our techniques to characterise photospheric granulation as an astrophysical noise source. A four component parameterisation of granulation is developed that can be used to reconstruct stellar line asymmetries and radial velocity shifts due to photospheric convective motions. The four components are made up of absorption line profiles calculated for granules, magnetic intergranular lanes, non-magnetic intergranular lanes, and magnetic bright points at disc centre. These components are constructed by averaging Fe I $6302 \mathrm{\AA}$ magnetically sensitive absorption line profiles output from detailed radiative transport calculations of the solar photosphere. Each of the four categories adopted are based on magnetic field and continuum intensity limits determined from examining three-dimensional magnetohydrodynamic simulations with an average magnetic flux of $200 \mathrm{G}$. Using these four component line profiles we accurately reconstruct granulation profiles, produced from modelling 12 x 12 Mm$^2$ areas on the solar surface, to within $\sim \pm$ 20 cm s$^{-1}$ on a $\sim$ 100 m s$^{-1}$ granulation signal. We have also successfully reconstructed granulation profiles from a $50 \mathrm{G}$ simulation using the parameterised line profiles from the $200 \mathrm{G}$ average magnetic field simulation. This test demonstrates applicability of the characterisation to a range of magnetic stellar activity levels.

Simulations of Biased Agonists in the β2 Adrenergic Receptor with Accelerated Molecular Dynamics

The biased agonism of the G protein-coupled receptors (GPCRs), where in addition to a traditional G protein-signalling pathway a GPCR promotes intracellular signals though ß-arrestin, is a novel paradigm in pharmacology. Biochemical and biophysical studies have suggested that a GPCR forms a distinct ensemble of conformations signalling through the G protein and ß-arrestin. Here we report on the dynamics of the ß2 adrenergic receptor bound to the ß-arrestin and G protein biased agonists and the empty receptor to further characterize the receptor conformational changes caused by biased agonists. We use conventional and accelerated molecular dynamics (aMD) simulations to explore the conformational transitions of the GPCR from the active state to the inactive state. We found that aMD simulations enable monitoring the transition within the nanosecond timescale while capturing the known microscopic characteristics of the inactive states, such as the ionic lock, the inward position of F6.44, and water clusters. Distinct conformational states are shown to be stabilized by each biased agonist. In particular, in simulations of the receptor with the ß-arrestin biased agonist, N-cyclopentylbutanepherine we observe a different pattern of motions in helix 7 when compared to simulations with the G protein biased agonist, Salbutamol that involves perturbations of the network of interactions within the NPxxY motif. Understanding the network of interactions induced by biased ligands and the subsequent receptor conformational shifts will lead to development of more efficient drugs. © 2013 American Chemical Society

Anticipatory Planning Reveals Segmentation of Cortical Motor Output During Action Observation

It has been argued that the variation in brain activity that occurs when observing another person reflects a representation of actions that is indivisible, and which plays out in full once the intent of the actor can be discerned. We used transcranial magnetic stimulation to probe the excitability of corticospinal projections to 2 intrinsic hand muscles while motions to reach and grasp an object were observed. A symbolic cue either faithfully indicated the required final orientation of the object and thus the nature of the grasp that was required, or was in conflict with the movement subsequently displayed. When the cue was veridical, modulation of excitability was in accordance with the functional role of the muscles in the action observed. If however the cue had indicated that the alternative grasp would be required, modulation of output to first dorsal interosseus was consistent with the action specified, rather than the action observed-until the terminal phase of the motion sequence during which the object was seen lifted. Modulation of corticospinal output during observation is thus segmented-it progresses initially in accordance with the action anticipated, and if discrepancies are revealed by visual input, coincides thereafter with that of the action seen.

Dynamic properties of bright points in an active region

Context. Bright points (BPs) are small-scale, magnetic features ubiquitous across the solar surface. Previously, we have observed and noted their properties for quiet Sun regions. Here, we determine the dynamic properties of BPs using simultaneous quiet Sun and active region data.

Aims. The aim of this paper is to compare the properties of BPs in both active and quiet Sun regions and to determine any difference in the dynamics and general properties of BPs as a result of the varying magnetic activity within these two regions.

Methods. High spatial and temporal resolution G-band observations of active region AR11372 were obtained with the Rapid Oscillations in the Solar Atmosphere instrument at the Dunn Solar Telescope. Three subfields of varying polarity and magnetic flux density were selected with the aid of magnetograms obtained from the Helioseismic and Magnetic Imager on board the Solar Dynamics Observatory. Bright points within these subfields were subsequently tracked and analysed.

Results. It is found that BPs within active regions display attenuated velocity distributions with an average horizontal velocity of ~0.6 km s-1, compared to the quiet region which had an average velocity of 0.9 km s-1. Active region BPs are also ~21% larger than quiet region BPs and have longer average lifetimes (~132 s) than their quiet region counterparts (88 s). No preferential flow directions are observed within the active region subfields. The diffusion index (γ) is estimated at ~1.2 for the three regions.

Conclusions. We confirm that the dynamic properties of BPs arise predominately from convective motions. The presence of stronger field strengths within active regions is the likely reason behind the varying properties observed. We believe that larger amounts of magnetic flux will attenuate BP velocities by a combination of restricting motion within the intergranular lanes and by increasing the number of stagnation points produced by inhibited convection. Larger BPs are found in regions of higher magnetic flux density and we believe that lifetimes increase in active regions as the magnetic flux stabilises the BPs.