In silico prediction of mutant HIV-1 proteases cleaving a target sequence

HIV-1 protease represents an appealing system for directed enzyme re-design, since it has various different endogenous targets, a relatively simple structure and it is well studied. Recently Chaudhury and Gray (Structure (2009) 17: 1636 -- 1648) published a computational algorithm to discern the specificity determining residues of HIV-1 protease. In this paper we present two computational tools aimed at re-designing HIV-1 protease, derived from the algorithm of Chaudhuri and Gray. First, we present an energy-only based methodology to discriminate cleavable and non cleavable peptides for HIV-1 proteases, both wild type and mutant. Secondly, we show an algorithm we developed to predict mutant HIV-1 proteases capable of cleaving a new target substrate peptide, different from the natural targets of HIV-1 protease. The obtained in silico mutant enzymes were analyzed in terms of cleavability and specificity towards the target peptide using the energy-only methodology. We found two mutant proteases as best candidates for specificity and cleavability towards the target sequence

Covariant EBK quantization of the electromagnetic two-body problem

We discuss a method to transform the covariant Fokker action into an implicit two-degree-of-freedom Hamiltonian for the electromagnetic two-body problem with arbitrary masses. This dynamical system appeared 100 years ago and it was popularized in the 1940's by the still incomplete Wheeler and Feynman program to quantize it as a means to overcome the divergencies of perturbative QED. Our finite-dimensional implicit Hamiltonian is closed and involves no series expansions. The Hamiltonian formalism is then used to motivate an EBK quantization based on the classical trajectories with a non-perturbative formula that predicts energies free of infinities.Comment: 21 page

Structural Properties of the Disordered Spherical and other Mean Field Spin Models

We extend the approach of Aizenman, Sims and Starr for the SK-type models to their spherical versions. Such an extension has already been performed for diluted spin glasses. The factorization property of the optimal structures found by Guerra for the SK model, which holds for diluted models as well, is verified also in the case of spherical systems, with the due modifications. Hence we show that there are some common structural features in various mean field spin models. These similarities seem to be quite paradigmatic, and we summarize the various techniques typically used to prove the structural analogies and to tackle the computation of the free energy per spin in the thermodynamic limit.Comment: 24 page

Long Range Magnetic Order and the Darwin Lagrangian

We simulate a finite system of $N$ confined electrons with inclusion of the Darwin magnetic interaction in two- and three-dimensions. The lowest energy states are located using the steepest descent quenching adapted for velocity dependent potentials. Below a critical density the ground state is a static Wigner lattice. For supercritical density the ground state has a non-zero kinetic energy. The critical density decreases with $N$ for exponential confinement but not for harmonic confinement. The lowest energy state also depends on the confinement and dimension: an antiferromagnetic cluster forms for harmonic confinement in two dimensions.Comment: 5 figure

A Computational Methodology to Screen Activities of Enzyme Variants

We present a fast computational method to efficiently screen enzyme activity. In the presented method, the effect of mutations on the barrier height of an enzyme-catalysed reaction can be computed within 24 hours on roughly 10 processors. The methodology is based on the PM6 and MOZYME methods as implemented in MOPAC2009, and is tested on the first step of the amide hydrolysis reaction catalyzed by Candida Antarctica lipase B (CalB) enzyme. The barrier heights are estimated using adiabatic mapping and are shown to give barrier heights to within 3kcal/mol of B3LYP/6-31G(d)//RHF/3-21G results for a small model system. Relatively strict convergence criteria (0.5kcal/(mol{\AA})), long NDDO cutoff distances within the MOZYME method (15{\AA}) and single point evaluations using conventional PM6 are needed for reliable results. The generation of mutant structure and subsequent setup of the semiempirical calculations are automated so that the effect on barrier heights can be estimated for hundreds of mutants in a matter of weeks using high performance computing

Fractional-Spin Integrals of Motion for the Boundary Sine-Gordon Model at the Free Fermion Point

We construct integrals of motion (IM) for the sine-Gordon model with boundary at the free Fermion point which correctly determine the boundary S matrix. The algebra of these IM (``boundary quantum group'' at q=1) is a one-parameter family of infinite-dimensional subalgebras of twisted affine sl(2). We also propose the structure of the fractional-spin IM away from the free Fermion point.Comment: 19 pages, LaTeX, no figure

Clustering of vertically constrained passive particles in homogeneous, isotropic turbulence

We analyze the dynamics of small particles vertically confined, by means of a linear restoring force, to move within a horizontal fluid slab in a three-dimensional (3D) homogeneous isotropic turbulent velocity field. The model that we introduce and study is possibly the simplest description for the dynamics of small aquatic organisms that, due to swimming, active regulation of their buoyancy, or any other mechanism, maintain themselves in a shallow horizontal layer below the free surface of oceans or lakes. By varying the strength of the restoring force, we are able to control the thickness of the fluid slab in which the particles can move. This allows us to analyze the statistical features of the system over a wide range of conditions going from a fully 3D incompressible flow (corresponding to the case of no confinement) to the extremely confined case corresponding to a two-dimensional slice. The background 3D turbulent velocity field is evolved by means of fully resolved direct numerical simulations. Whenever some level of vertical confinement is present, the particle trajectories deviate from that of fluid tracers and the particles experience an effectively compressible velocity field. Here, we have quantified the compressibility, the preferential concentration of the particles, and the correlation dimension by changing the strength of the restoring force. The main result is that there exists a particular value of the force constant, corresponding to a mean slab depth approximately equal to a few times the Kolmogorov length scale, that maximizes the clustering of the particles

The way to ultrafast, high-throughput enantioseparations of bioactive compounds in liquid and supercritical fluid chromatography

Until less than 10 years ago, chiral separations were carried out with columns packed with 5 or 3 μm fully porous particles (FPPs). Times to resolve enantiomeric mixtures were easily larger than 30 min, or so. Pushed especially by stringent requirements from medicinal and pharmaceutical industries, during the last years the field of chiral separations by liquid chromatography has undergone what can be defined a “true revolution”. With the purpose of developing ever faster and efficient method of separations, indeed, very efficient particle formats, such as superficially porous particles (SPPs) or sub-2 μm FPPs, have been functionalized with chiral selectors and employed in ultrafast applications. Thanks to the use of short column (1–2 cm long), packed with these extremely efficient chiral stationary phases (CSPs), operated at very high flow rates (5–8 mL/min), resolution of racemates could be accomplished in very short time, in many cases less than 1 s in normal-, reversed-phase and HILIC conditions. These CSPs have been found to be particularly promising also to carry out high-throughput separations under supercritical fluid chromatography (SFC) conditions. The most important results that have been recently achieved in terms of ultrafast, high-throughput enantioseparations both in liquid and supercritical fluid chromatography with particular attention to the very important field of bioactive chiral compounds will be reviewed in this manuscript. Attention will be focused not only on the latest introduced CSPs and their applications, but also on instrumental modifications which are required in some cases in order to fully exploit the intrinsic potential of new generation chiral columns

Decision Problems For Convex Languages

In this paper we examine decision problems associated with various classes of convex languages, studied by Ang and Brzozowski (under the name "continuous languages"). We show that we can decide whether a given language L is prefix-, suffix-, factor-, or subword-convex in polynomial time if L is represented by a DFA, but that the problem is PSPACE-hard if L is represented by an NFA. In the case that a regular language is not convex, we prove tight upper bounds on the length of the shortest words demonstrating this fact, in terms of the number of states of an accepting DFA. Similar results are proved for some subclasses of convex languages: the prefix-, suffix-, factor-, and subword-closed languages, and the prefix-, suffix-, factor-, and subword-free languages.Comment: preliminary version. This version corrected one typo in Section 2.1.1, line