Crystallization and preliminary X-ray diffraction analysis of three myotoxic phospholipases A2 from Bothrops brazili venom

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Molecular detection of enterotoxins E, G, H and I in Staphylococcus aureus and coagulase-negative staphylococci isolated from clinical samples of newborns in Brazil

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Docking and small angle X-ray scattering studies of purine nucleoside phosphorylase

Docking simulations have been used to assess protein complexes with some success. Small angle X-ray scattering (SAXS) is a well-established technique to investigate protein spatial configuration. This work describes the integration of geometric docking with SAXS to investigate the quaternary structure of recombinant human purine nucleoside phosphorylase (PNP). This enzyme catalyzes the reversible phosphorolysis of N-ribosidic bonds of purine nucleosides and deoxynucleosides. A genetic deficiency due to mutations in the gene encoding for PNP causes gradual decrease in T-cell immunity. Inappropriate activation of T-cells has been implicated in several clinically relevant human conditions such as transplant rejection, rheumatoid arthritis, lupus, and T-cell lymphomas. PNP is therefore a target for inhibitor development aiming at T-cell immune response modulation and has been submitted to extensive structure-based drug design. The present analysis confirms the trimeric structure observed in the crystal. The potential application of the present procedure to other systems is discussed. (C) 2003 Elsevier B.V. All rights reserved.

Assignment of chromosome rearrangements between X chromosomes of human and cattle by laser microdissection and Zoo-FISH

Cross-species fluorescence in-situ hybridization (Zoo-FISH) was performed on cattle metaphase spreads using Homo sapiens X chromosome (HSAX) painting probes specific for the p- and q-arms to identify the cytogenetic location of a chromosome breakpoint between HSAX and the Bos taurus X chromosome (BTAX). The existence of a breakpoint is strongly suggested by recent radiation hybrid and FISH mapping results. Hybridization probes were generated by microdissection of HSAX p- and q-arms using the contact-free technology of Laser Microdissection and Pressure Catapulting (LMPC), amplification of the isolated chromosome material by DOP-PCR, and labelling of the PCR products with digoxigenin in a secondary PCR. Independent Zoo-FISH of the two painting probes on bovine metaphase chromosomes (detected by antidigoxigenin-fluorescein) resulted in clear hybridization signals on BTAX. A breakpoint was identified between HSAXp and HSAXq on BTAX, and narrowed down between the G-bands BTAXq25 and BTAXq26. The assumed centromere transposition between HSAX and BTAX associated with the rearranged chromosome segments is supported by cytogenetic assignments of the genes BGN and G6PD to BTAX.

Crystallization and preliminary X-ray analysis of bucain, a novel toxin from the Malayan krait Bungarus candidus

Bucain is a three-finger toxin, structurally homologous to snake-venom muscarinic toxins, from the venom of the Malayan krait Bungarus candidus. These proteins have molecular masses of approximately 6000-8000 da and encompass the potent curaremimetic neurotoxins which confer lethality to Elapidae and Hydrophidae venoms. Bucain was crystallized in two crystal forms by the hanging-drop vapour-diffusion technique in 0.1 M sodium citrate pH 5.6, 15% PEG 4000 and 0.15 M ammonium acetate. Form I crystals belong to the monoclinic system space group C2, with unit-cell parameters a = 93.73, b = 49.02, c = 74.09 Angstrom, beta = 111.32degrees, and diffract to a nominal resolution of 1.61 Angstrom. Form II crystals also belong to the space group C2, with unit-cell parameters a = 165.04, b = 49.44, c = 127.60 Angstrom, beta = 125.55degrees, and diffract to a nominal resolution of 2.78 Angstrom. The self-rotation function indicates the presence of four and eight molecules in the crystallographic asymmetric unit of the form I and form II crystals, respectively. Attempts to solve these structures by molecular-replacement methods have not been successful and a heavy-atom derivative search has been initiated.

Thermal and Structural Behavior of Dioctadecyldimethylammonium Bromide Dispersions Studied by Differential Scanning Calorimetry and X-Ray Scattering

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Crystallization, preliminary X-ray diffraction analysis and Patterson search of oxyhaemoglobin I from the wolf (Chrysocyon brachiurus)

Oxyhaemoglobin I isolated from the Brazilian wolf Chrysocyon brachiurus has been crystallized and X-ray diffraction data has been collected to 2.06 A resolution using a synchrotron-radiation source. Crystals were determined to belong to the space group P2(1)2(1)2(1) and preliminary structural analysis revealed the presence of one tetramer in the asymmetric unit. The structure was determined using standard molecular-replacement techniques and is currently being refined using maximum-likelihood protocols. This is the first haemoglobin isolated from a member of the Canidae family to be crystallized and it will provide further insights in the comparative biochemistry of vertebrate haemoglobins.

Measurement of the p(p)over-bar -> WZ+X cross section at root s=1.96 TeV and limits on WWZ trilinear gauge couplings

We present measurements of the process p (P) over bar -> WZ + X -> l 'nu(l ')l (l) over bar at root s = 1:96 TeV,where l and l ' are electrons or muons. Using 1 fb(-1) of data from the D0 experiment, we observe 13 candidates with an expected background of 4.5 +/- 0.6 events and measure a cross section sigma(WZ) = 2.7(-1.3)(+1.7) pb. From the number of observed events and the Z boson transverse momentum distribution, we limit the trilinear WWZ gauge couplings to -0: 17 <= lambda(Z) <= 0.21 (Delta k(Z) <= 0.29(lambda(Z) = 0) at the 95% C.L. for a form factor scale Lambda = 2 TeV. Further, assuming that Delta g(1)(Z) = Delta k(Z), we find -0.12 <= Delta k(Z) <= 0.29(lambda(Z) = 0) at the 95% C. L. These are the most restrictive limits on the WWZ couplings available to date.

Measurement of the p(p)over-bar -> t(t)over-bar + X production cross section at root s=1.96 TeV in the fully hadronic decay channel

A measurement of the top quark pair production cross section in proton antiproton collisions at an interaction energy of root s=1.96 TeV is presented. This analysis uses 405 +/- 25 pb(-1) of data collected with the D0 detector at the Fermilab Tevatron Collider. Fully hadronic t (t) over bar decays with final states of six or more jets are separated from the multijet background using secondary vertex tagging and a neural network. The t (t) over bar cross section is measured as sigma(t (t) over bar)=4.5(-1.9)(+2.0)(stat)(-1.1)(+1.4)(syst)+/- 0.3(lumi) pb for a top quark mass of m(t)=175 GeV/c(2).

Search for associated Higgs boson production WH -> WWW*-> l(+/-)nu l('+/-)nu(')+X in p(p)over-bar collisions at root S=1.96 TeV

We present a search for associated Higgs boson production in the process p (p) over bar -> WH -> WWW*-> l(+/-)nu l('+/-)nu(')+X in final states containing two like-sign isolated electrons or muons (e(+/-)e(+/-), e(+/-)mu(+/-), or mu(+/-)mu(+/-)). The search is based on D0 run II data samples corresponding to integrated luminosities of 360-380 pb(-1). No excess is observed over the predicted standard model background. We set 95% C.L. upper limits on sigma ->(p (p) over bar WH) x Br(H -> WW*) between 3.2 and 2.8 pb for Higgs boson masses from 115 to 175 GeV.

Measurement of the shape of the boson rapidity distribution for p(p)over-bar -> Z/gamma(*)-> e(+)e(-)+X events produced at root s of 1.96 TeV

We present a measurement of the shape of the boson rapidity distribution for p (p) over bar -> Z/gamma(*)-> e(+)e(-)+X events at a center-of-mass energy of 1.96 TeV. The measurement is made for events with electron-positron mass 71 < M-ee < 111 GeV and uses 0.4 fb(-1) of data collected at the Fermilab Tevatron collider with the D0 detector. This measurement significantly reduces the uncertainties on the rapidity distribution in the forward region compared with previous measurements. Predictions of next-to-next-to-leading order (NNLO) QCD are found to agree well with the data over the full rapidity range.

Observation and properties of the X(3872) decaying to J/psi pi(+)pi(-) in p(p)over-bar collisions at root s=1.96 TeV

We report the observation of the X(3872) in the J/psipi(+)pi(-) channel, with J/psi decaying to mu(+)mu(-), in p (p) over bar collisions at roots=1.96 TeV. Using approximately 230 pb(-1) of data collected with the Run II D0 detector, we observe 522+/-100 X(3872) candidates. The mass difference between the X(3872) state and the J/psi is measured to be 774.9+/-3.1(stat)+/-3.0(syst) MeV/c(2). We have investigated the production and decay characteristics of the X(3872) and find them to be similar to those of the psi(2S) state.

Measurement of the CP-violation parameter of B-0 mixing and decay with p(p)over-bar ->mu mu X data

We measure the dimuon charge asymmetry A in p (p) over bar collisions at a center of mass energy root s=1960 GeV. The data was recorded with the D0 detector and corresponds to an integrated luminosity of approximately 1.0 fb(-1). Assuming that the asymmetry A is due to asymmetric B-0 <->(B) over bar (0) mixing and decay, we extract the CP-violation parameter of B-0 mixing and decay: ((epsilon B0))/(1+vertical bar epsilon B0 vertical bar 2)=(AB0)/(4)= -0.0023 +/- 0.0011(stat)+/- 0.0008(syst).A(B)(0) is the dimuon charge asymmetry from decays of B-0(B) over bar (0) pairs. The general case, with CP violation in both B-0 and B-s(0) systems, is also considered. Finally we obtain the forward-backward asymmetry that quantifies the tendency of mu(+) to go in the proton direction and mu(-) to go in the antiproton direction. The results are consistent with the standard model and constrain new physics.

Measurement of the pp(-)-> W gamma plus X cross section at root s=1.96 TeV and WW gamma anomalous coupling limits

The WW gamma triple gauge boson coupling parameters are studied using p (p) over bar -> l nu gamma + X(l = e, mu) events at root s = 1.96 TeV. The data were collected with the D0 detector from an integrated luminosity of 162 pb(-1) delivered by the Fermilab Tevatron Collider. The cross section times branching fraction for p (p) over bar -> W(gamma) + X -> l nu gamma + X with E-T(gamma) > 8 GeV and Delta R-l gamma > 0.7 is 14.8 +/- 1.6(stat) +/- 1.0(syst) +/- 1.0(lum) pb. The one-dimensional 95% confidence level limits on anomalous couplings are -0.88 < Delta kappa(gamma) < 0.96 and -0.20 < lambda(gamma) < 0.20.

Measurement of sigma(p(p)over-bar -> Z plus X)Br(Z -> tau(+)tau(-)) at root s=1.96 TeV

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)