Extensive Molecular Dynamics Simulations Disclosed the Stability of mPGES-1 Enzyme and the Structural Role of Glutathione (GSH) Cofactor

A deep in silico investigation of various microsomal prostaglandin E-2 synthase-1 (mPGES-1) protein systems is here reported using molecular dynamics (MD) simulations. Firstly, eight different proteins models (Models A-H) were built, starting from the active enzyme trimer system (Model A), namely that bound to three glutathione (GSH) cofactor molecules, and then gradually removing the GSHs (Models B-H), simulating each of them for 100 ns in explicit solvent. The analysis of the obtained data disclosed the structural role of GSH in the chemical architecture of mPGES-1 enzyme, thus suggesting the unlikely displacement of this cofactor, in accordance with experimentally determined protein structures co-complexed with small molecule inhibitors. Afterwards, Model A was submitted to microsecond-scale molecular dynamics simulation (total simulation time=10 mu s), in order to shed light about the dynamical behaviour of this enzyme at atomic level and to obtain further structural features and protein function information. We confirmed the structural stability of the enzyme machinery, observing a conformational rigidity of the protein, with a backbone RMSD of similar to 3 angstrom along the simulation time, and highlighting the strong active contribution of GSH molecules due to their active role in packing the protein chains through a tight binding at monomer interfaces. Furthermore, the focused analysis on R73 residue disclosed its role in solvent exchange events, probably excluding its function as route for GSH to enter towards the endoplasmic reticulum membrane, in line with the recently reported function of cap domain residues F44-D66 as gatekeeper for GSH entrance into catalytic site

Plakilactones G and H from a marine sponge. Stereochemical determination of highly flexible systems by quantitative NMR-derived interproton distances combined with quantum mechanical calculations of C-13 chemical shifts

In this paper the stereostructural investigation of two new oxygenated polyketides, plakilactones G and H, isolated from the marine sponge Plakinastrella mamillaris collected at Fiji Islands, is reported. The stereostructural studies began on plakilactone H by applying an integrated approach of the NOE-based protocol and quantum mechanical calculations of (13)C chemical shifts. In particular, plakilactone H was used as a template to extend the application of NMR-derived interproton distances to a highly flexible molecular system with simultaneous assignment of four non-contiguous stereocenters. Chemical derivatization and quantum mechanical calculations of (13)C on plakilactone G along with a plausible biogenetic interconversion between plakilactone G and plakilactone H allowed us to determine the absolute configuration in this two new oxygenated polyketides

Overcome Chemoresistance: Biophysical and Structural Analysis of Synthetic FHIT-Derived Peptides

The fragile histidine triad (FHIT) protein is a member of the large and ubiquitous histidine triad (HIT) family of proteins. On the basis of genetic evidence, it has been postulated that the FHIT protein may function as tumor suppressor, implying a role for the FHIT protein in carcinogenesis. Recently, Gaudio et al. reported that FHIT binds and delocalizes annexin A4 (ANXA4) from plasma membrane to cytosol in paclitaxel-resistant lung cancer cells, thus restoring their chemosensitivity to the drug. They also identified the smallest protein sequence of the FHIT still interacting with ANXA4, ranging fromposition 7 to 13: QHLIKPS. This short sequence of FHIT protein was not only able to bind ANXA4 but also to hold its target in the cytosol during paclitaxel treatment, thus avoiding ANXA4 translocation to the inner side of the cell membrane. Starting from these results, to obtain much information about structure requirements involved in the interaction of the peptide mentioned above, we synthetized a panel of seven peptides through an Ala-scan approach. In detail, to study the binding of FHIT derived peptides with ANXA4, we applied a combination of different biophysical techniques such as differential scanning fluorimetry (DSF), surface plasmon resonance (SPR), and microscale thermophoresis (MST). Circular dichroism (CD) and nuclear magnetic resonance (NMR) were used to determine the conformational structure of the lead peptide (7–13) and peptides generated from ala-scan technique. The application of different biophysical and structural techniques, integrated by a preliminary biological evaluation, allowed us to build a solid structure activity relationship on the synthesized peptides

Identification of 2,4-Dinitro-Biphenyl-Based Compounds as MAPEG Inhibitors

We identified 2,4-dinitro-biphenyl-based compounds as new inhibitors of leukotriene C-4 synthase (LTC4S) and 5-lipoxygenase-activating protein (FLAP), both members of the "Membrane Associated Proteins in Eicosanoid and Glutathione metabolism" (MAPEG) family involved in the biosynthesis of pro-inflammatory eicosanoids. By molecular docking we evaluated the putative binding against the targets of interest, and by applying cell-free and cell-based assays we assessed the inhibition of LTC4S and FLAP by the small molecules at low micromolar concentrations. The present results integrate the previously observed inhibitory profile of the tested compounds against another MAPEG member, i. e., microsomal prostaglandin E-2 synthase (mPGES)-1, suggesting that the 2,4-dinitro-biphenyl scaffold is a suitable molecular platform for a multitargeting approach to modulate pro-inflammatory mediators in inflammation and cancer treatment

A global fit to determine the pseudoscalar mixing angle and the gluonium content of the eta' meson

We update the values of the eta-eta' mixing angle and of the eta' gluonium content by fitting our measurement R_phi = BR(phi to eta' gamma)/ BR(phi to eta gamma) together with several vector meson radiative decays to pseudoscalars (V to P gamma), pseudoscalar mesons radiative decays to vectors (P to V gamma) and the eta' to gamma gamma, pi^0 to gamma gamma widths. From the fit we extract a gluonium fraction of Z^2_G = 0.12 +- 0.04, the pseudoscalar mixing angle psi_P = (40.4 +- 0.6) degree and the phi-omega mixing angle psi_V = (3.32 +- 0.09) degree. Z^2_G and psi_P are fairly consistent with those previously published. We also evaluate the impact on the eta' gluonium content determination of future experimental improvements of the eta' branching ratios and decay width.Comment: 13 pages, 7 figures to submit to JHE

Precise measurements of the eta and the neutral kaon meson masses with the KLOE detector

We present precise measurements of the eta and K0 masses using the processes phi to eta gamma, eta to gamma gamma and phi to Ks Kl, Ks to pi+ pi-. The K0 mass measurement, M_K=497.583 +/- 0.005 (stat) +/- 0.020 (syst) MeV, is in acceptable agreement with the previous measurements but is more accurate. We find m(eta) = 547.874 +/- 0.007 (stat) +/- 0.031 (syst) MeV. Our value is the most accurate to date and is in agreement with two recent measurements based on eta decays, but is inconsistent, by about 10 sigma, with a measurement of comparable precision based on eta production at threshold.Comment: 15 pages, 8 figures Submitted to Physics Letters

Study of the process e+e- -> omega pi0 in the phi-meson mass region with the KLOE detector

We have studied the e+e- -> omegapi0 cross section in the sqrt(s) interval 1000-1030 MeV using the pi+pi-pi0pi0 and pi0pi0gamma final states with a sample of ~600 pb^-1 collected with the KLOE detector at DAFNE. By fitting the observed interference pattern around M_phi for both final states, we extract the ratio of the decay widths Gamma(omega->pi0gamma)/Gamma(omega->pi+pi-pi0) = 0.0897 +- 0.0016 and derive the branching fractions BR(omega -> pi+pi-pi0)= (90.24 +- 0.19)%, BR(omega -> pi0gamma) = (8.09 +- 0.14)%. The parameters describing the e+e- -> omegapi0 reaction around M_\phi are also used to extract the branching fraction for the OZI and G-parity violating phi -> omegapi0 decay: BR(phi->omegapi0) = (4.4 +- 0.6)x10^-5.Comment: 12 Pages, 4 figures, submitted to Physics Letter

Search for the K_S -> e+e- decay with the KLOE detector

We present the result of a direct search for the decay K_S -> e+e-, obtained with a sample of e+e- -> phi -> K_S K_L events produced at DAFNE, the Frascati phi-factory, for an integrated luminosity of 1.9 fb^-1. The search has been performed using a pure K_S beam tagged by the simultaneous detection of a K_L interaction in the calorimeter. Background rejection has been optimized by using both kinematic and particle identification cuts. We find BR(K_S -> e+e-) < 9x10^-9 at 90% CL, which improves by an order of magnitude on the previous best limit.Comment: 13 pages, 11 figures, submitted to Physics Letters

Charged kaon lifetime at KLOE

Preliminary result on the charged kaon lifetime, obtained by the KLOE experiment operating at DA$\Phi$NE, the Frascati $\phi$-factory, is presentedComment: 3 pages, 3 figures, to appear in the proceedings of 42nd Rencontres de Moriond on Electroweak Interactions and Unified Theories, La Thuile, Aosta Valley, Italy, 10-17 Mar 200