707 research outputs found

    Mechanism of Protein Kinetic Stabilization by Engineered Disulfide Crosslinks

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    The impact of disulfide bonds on protein stability goes beyond simple equilibrium thermodynamics effects associated with the conformational entropy of the unfolded state. Indeed, disulfide crosslinks may play a role in the prevention of dysfunctional association and strongly affect the rates of irreversible enzyme inactivation, highly relevant in biotechnological applications. While these kinetic-stability effects remain poorly understood, by analogy with proposed mechanisms for processes of protein aggregation and fibrillogenesis, we propose that they may be determined by the properties of sparsely-populated, partially-unfolded intermediates. Here we report the successful design, on the basis of high temperature molecular-dynamics simulations, of six thermodynamically and kinetically stabilized variants of phytase from Citrobacter braakii (a biotechnologically important enzyme) with one, two or three engineered disulfides. Activity measurements and 3D crystal structure determination demonstrate that the engineered crosslinks do not cause dramatic alterations in the native structure. The inactivation kinetics for all the variants displays a strongly non-Arrhenius temperature dependence, with the time-scale for the irreversible denaturation process reaching a minimum at a given temperature within the range of the denaturation transition. We show this striking feature to be a signature of a key role played by a partially unfolded, intermediate state/ensemble. Energetic and mutational analyses confirm that the intermediate is highly unfolded (akin to a proposed critical intermediate in the misfolding of the prion protein), a result that explains the observed kinetic stabilization. Our results provide a rationale for the kinetic-stability consequences of disulfide-crosslink engineering and an experimental methodology to arrive at energetic/structural descriptions of the sparsely populated and elusive intermediates that play key roles in irreversible protein denaturation.This work was supported by grants BIO2009-09562, CSD2009-00088 from the Spanish Ministry of Science and Innovation, and FEDER Funds (JMS-R)

    Measurement of the Dipion Mass Spectrum in X(3872) -> J/Psi Pi+ Pi- Decays

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    We measure the dipion mass spectrum in X(3872)--> J/Psi Pi+ Pi- decays using 360 pb-1 of pbar-p collisions at 1.96 TeV collected with the CDF II detector. The spectrum is fit with predictions for odd C-parity (3S1, 1P1, and 3DJ) charmonia decaying to J/Psi Pi+ Pi-, as well as even C-parity states in which the pions are from Rho0 decay. The latter case also encompasses exotic interpretations, such as a D0-D*0Bar molecule. Only the 3S1 and J/Psi Rho hypotheses are compatible with our data. Since 3S1 is untenable on other grounds, decay via J/Psi Rho is favored, which implies C=+1 for the X(3872). Models for different J/Psi-Rho angular momenta L are considered. Flexibility in the models, especially the introduction of Rho-Omega interference, enable good descriptions of our data for both L=0 and 1.Comment: 7 pages, 4 figures -- Submitted to Phys. Rev. Let

    Precision measurement of the top quark mass from dilepton events at CDF II

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    We report a measurement of the top quark mass, M_t, in the dilepton decay channel of ttˉb+νbˉνˉt\bar{t}\to b\ell'^{+}\nu_{\ell'}\bar{b}\ell^{-}\bar{\nu}_{\ell} using an integrated luminosity of 1.0 fb^{-1} of p\bar{p} collisions collected with the CDF II detector. We apply a method that convolutes a leading-order matrix element with detector resolution functions to form event-by-event likelihoods; we have enhanced the leading-order description to describe the effects of initial-state radiation. The joint likelihood is the product of the likelihoods from 78 candidate events in this sample, which yields a measurement of M_{t} = 164.5 \pm 3.9(\textrm{stat.}) \pm 3.9(\textrm{syst.}) \mathrm{GeV}/c^2, the most precise measurement of M_t in the dilepton channel.Comment: 7 pages, 2 figures, version includes changes made prior to publication by journa

    Measurement of the Ratios of Branching Fractions B(Bs -> Ds pi pi pi) / B(Bd -> Dd pi pi pi) and B(Bs -> Ds pi) / B(Bd -> Dd pi)

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    Using 355 pb^-1 of data collected by the CDF II detector in \ppbar collisions at sqrt{s} = 1.96 TeV at the Fermilab Tevatron, we study the fully reconstructed hadronic decays B -> D pi and B -> D pi pi pi. We present the first measurement of the ratio of branching fractions B(Bs -> Ds pi pi pi) / B(Bd -> Dd pi pi pi) = 1.05 pm 0.10 (stat) pm 0.22 (syst). We also update our measurement of B(Bs -> Ds pi) / B(Bd -> Dd pi) to 1.13 pm 0.08 (stat) pm 0.23 (syst) improving the statistical uncertainty by more than a factor of two. We find B(Bs -> Ds pi) = [3.8 pm 0.3 (stat) pm 1.3 (syst)] \times 10^{-3} and B(Bs -> Ds pi pi pi) = [8.4 pm 0.8 (stat) pm 3.2 (syst)] \times 10^{-3}.Comment: 7 pages, 2 figure

    Cross Section Measurements of High-pTp_T Dilepton Final-State Processes Using a Global Fitting Method

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    We present a new method for studying high-pTp_T dilepton events (e±ee^{\pm}e^{\mp}, μ±μ\mu^{\pm}\mu^{\mp}, e±μe^{\pm}\mu^{\mp}) and simultaneously extracting the production cross sections of ppˉttˉp\bar{p} \to t\bar{t}, ppˉW+Wp\bar{p} \to W^+W^-, and p\bar{p} \to \ztt at a center-of-mass energy of s=1.96\sqrt{s} = 1.96 TeV. We perform a likelihood fit to the dilepton data in a parameter space defined by the missing transverse energy and the number of jets in the event. Our results, which use 360pb1360 {\rm pb^{-1}} of data recorded with the CDF II detector at the Fermilab Tevatron Collider, are σ(ttˉ)=8.52.2+2.7\sigma(t\bar{t}) = 8.5_{-2.2}^{+2.7} pb, σ(W+W)=16.34.4+5.2\sigma(W^+W^-) = 16.3^{+5.2}_{-4.4} pb, and \sigma(\ztt) =291^{+50}_{-46} pb.Comment: 20 pages, 2 figures, to be submitted to PRD-R

    Top Quark Mass Measurement from Dilepton Events at CDF II with the Matrix-Element Method

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    We describe a measurement of the top quark mass using events with two charged leptons collected by the CDF II detector from ppˉp\bar{p} collisions with s=1.96\sqrt s = 1.96 TeV at the Fermilab Tevatron. The likelihood in top mass is calculated for each event by convoluting the leading order matrix element describing qqˉttˉbνbˉνq\bar{q} \to t\bar{t} \to b\ell\nu_{\ell}\bar{b}\ell'\nu_{\ell'} with detector resolution functions. The presence of background events in the data sample is modeled using similar calculations involving the matrix elements for major background processes. In a data sample with integrated luminosity of 340 pb1^{-1}, we observe 33 candidate events and measure Mtop=165.2±6.1(stat.)±3.4(syst.) GeV/c2.M_{top} = 165.2 \pm 6.1(\textrm{stat.}) \pm 3.4(\textrm{syst.}) \mathrm{~GeV}/c^2. This measurement represents the first application of this method to events with two charged leptons and is the most precise single measurement of the top quark mass in this channel.Comment: 21 pages, 14 figure
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