401 research outputs found
Expression in E. coli and characterization of the catalytic domain of Botrytis cinerea chitin synthase
<p>Abstract</p> <p>Background</p> <p>Chitin synthase 3a (CHS3a) from <it>Botrytis cinerea </it>(Bc) catalyses the multiple transfer of <it>N</it>-acetylglucosamine (GlcNAc) residues to the growing chitin chain. Chitin, a β-1,4 linked GlcNAc homopolymer, is an essential cell wall component of filamentous fungi. Chitin synthase, processive membranous protein, has been recognized as a promising target for new antifungicides. Enzymatic characterizations of chitin synthases have been limited, mainly because purity and amounts of integral enzyme obtained after purification procedures have not been sufficient.</p> <p>Findings</p> <p>We undertook the preparation of two BcCHS3a fragment proteins, containing only the central domain and devoid of the N-terminal and transmembrane C-terminal regions. The central domain of CHS3a, named SGC (Spsa GntI Core), is conserved in all UDP-glycosyltransferases and it is believed to contain the active site of the enzyme. CHS3a-SGC protein was totally expressed as inclusion bodies in <it>Escherichia coli</it>. We performed recombinant CHS3a-SGC purification in denaturing conditions, followed by a refolding step. Although circular dichroism spectra clearly exhibited secondary structures of renatured CHS3a-SGC, no chitin synthase activity was detected. Nevertheless CHS3a-SGC proteins show specific binding for the substrate UDP-GlcNAc with a dissociation constant similar to the Michaelis constant and a major contribution of the uracil moiety for recognition was confirmed.</p> <p>Conclusions</p> <p>Milligram-scale quantities of CHS3a-SGC protein with native-like properties such as specific substrate UDP-GlcNAc binding could be easily obtained. These results are encouraging for subsequent heterologous expression of full-length CHS3a.</p
Saccharomyces cerevisiae chitin biosynthesis activation by N-acetylchitooses depends on size and structure of chito-oligosaccharides
<p>Abstract</p> <p>Background</p> <p>To explore chitin synthesis initiation, the effect of addition of exogenous oligosaccharides on <it>in vitro </it>chitin synthesis was studied. Oligosaccharides of various natures and lengths were added to a chitin synthase assay performed on a <it>Saccharomyces cerevisiae </it>membrane fraction.</p> <p>Findings</p> <p><it>N</it>-acetylchito-tetra, -penta and -octaoses resulted in 11 to 25% [<sup>14</sup>C]-GlcNAc incorporation into [<sup>14</sup>C]-chitin, corresponding to an increase in the initial velocity. The activation appeared specific to <it>N</it>-acetylchitooses as it was not observed with oligosaccharides in other series, such as beta-(1,4), beta-(1,3) or alpha-(1,6) glucooligosaccharides.</p> <p>Conclusions</p> <p>The effect induced by the <it>N</it>-acetylchitooses was a saturable phenomenon and did not interfere with free GlcNAc and trypsin which are two known activators of yeast chitin synthase activity <it>in vitro</it>. The magnitude of the activation was dependent on both oligosaccharide concentration and oligosaccharide size.</p
The Peculiar SN 2005hk: Do Some Type Ia Supernovae Explode as Deflagrations?
We present extensive u'g'r'i'BVRIYJHKs photometry and optical spectroscopy of
SN 2005hk. These data reveal that SN 2005hk was nearly identical in its
observed properties to SN 2002cx, which has been called ``the most peculiar
known type Ia supernova.'' Both supernovae exhibited high ionization SN
1991T-like pre-maximum spectra, yet low peak luminosities like SN 1991bg. The
spectra reveal that SN 2005hk, like SN 2002cx, exhibited expansion velocities
that were roughly half those of typical type Ia supernovae. The R and I light
curves of both supernovae were also peculiar in not displaying the secondary
maximum observed for normal type Ia supernovae. Our YJH photometry of SN 2005hk
reveals the same peculiarity in the near-infrared. By combining our optical and
near-infrared photometry of SN 2005hk with published ultraviolet light curves
obtained with the Swift satellite, we are able to construct a bolometric light
curve from ~10 days before to ~60 days after B maximum. The shape and unusually
low peak luminosity of this light curve, plus the low expansion velocities and
absence of a secondary maximum at red and near-infrared wavelengths, are all in
reasonable agreement with model calculations of a 3D deflagration which
produces ~0.25 M_sun of 56Ni.Comment: Accepted by PASP, to appear in April 2007 issue, 63 pages, 16
figures, 11 table
Muon Track Reconstruction and Data Selection Techniques in AMANDA
The Antarctic Muon And Neutrino Detector Array (AMANDA) is a high-energy
neutrino telescope operating at the geographic South Pole. It is a lattice of
photo-multiplier tubes buried deep in the polar ice between 1500m and 2000m.
The primary goal of this detector is to discover astrophysical sources of high
energy neutrinos. A high-energy muon neutrino coming through the earth from the
Northern Hemisphere can be identified by the secondary muon moving upward
through the detector. The muon tracks are reconstructed with a maximum
likelihood method. It models the arrival times and amplitudes of Cherenkov
photons registered by the photo-multipliers. This paper describes the different
methods of reconstruction, which have been successfully implemented within
AMANDA. Strategies for optimizing the reconstruction performance and rejecting
background are presented. For a typical analysis procedure the direction of
tracks are reconstructed with about 2 degree accuracy.Comment: 40 pages, 16 Postscript figures, uses elsart.st
On the selection of AGN neutrino source candidates for a source stacking analysis with neutrino telescopes
The sensitivity of a search for sources of TeV neutrinos can be improved by
grouping potential sources together into generic classes in a procedure that is
known as source stacking. In this paper, we define catalogs of Active Galactic
Nuclei (AGN) and use them to perform a source stacking analysis. The grouping
of AGN into classes is done in two steps: first, AGN classes are defined, then,
sources to be stacked are selected assuming that a potential neutrino flux is
linearly correlated with the photon luminosity in a certain energy band (radio,
IR, optical, keV, GeV, TeV). Lacking any secure detailed knowledge on neutrino
production in AGN, this correlation is motivated by hadronic AGN models, as
briefly reviewed in this paper.
The source stacking search for neutrinos from generic AGN classes is
illustrated using the data collected by the AMANDA-II high energy neutrino
detector during the year 2000. No significant excess for any of the suggested
groups was found.Comment: 43 pages, 12 figures, accepted by Astroparticle Physic
Calibration and Characterization of the IceCube Photomultiplier Tube
Over 5,000 PMTs are being deployed at the South Pole to compose the IceCube
neutrino observatory. Many are placed deep in the ice to detect Cherenkov light
emitted by the products of high-energy neutrino interactions, and others are
frozen into tanks on the surface to detect particles from atmospheric cosmic
ray showers. IceCube is using the 10-inch diameter R7081-02 made by Hamamatsu
Photonics. This paper describes the laboratory characterization and calibration
of these PMTs before deployment. PMTs were illuminated with pulses ranging from
single photons to saturation level. Parameterizations are given for the single
photoelectron charge spectrum and the saturation behavior. Time resolution,
late pulses and afterpulses are characterized. Because the PMTs are relatively
large, the cathode sensitivity uniformity was measured. The absolute photon
detection efficiency was calibrated using Rayleigh-scattered photons from a
nitrogen laser. Measured characteristics are discussed in the context of their
relevance to IceCube event reconstruction and simulation efforts.Comment: 40 pages, 12 figure
Limits on the high-energy gamma and neutrino fluxes from the SGR 1806-20 giant flare of December 27th, 2004 with the AMANDA-II detector
On December 27th 2004, a giant gamma flare from the Soft Gamma-ray Repeater
1806-20 saturated many satellite gamma-ray detectors. This event was by more
than two orders of magnitude the brightest cosmic transient ever observed. If
the gamma emission extends up to TeV energies with a hard power law energy
spectrum, photo-produced muons could be observed in surface and underground
arrays. Moreover, high-energy neutrinos could have been produced during the SGR
giant flare if there were substantial baryonic outflow from the magnetar. These
high-energy neutrinos would have also produced muons in an underground array.
AMANDA-II was used to search for downgoing muons indicative of high-energy
gammas and/or neutrinos. The data revealed no significant signal. The upper
limit on the gamma flux at 90% CL is dN/dE < 0.05 (0.5) TeV^-1 m^-2 s^-1 for
gamma=-1.47 (-2). Similarly, we set limits on the normalization constant of the
high-energy neutrino emission of 0.4 (6.1) TeV^-1 m^-2 s^-1 for gamma=-1.47
(-2).Comment: 14 pages, 3 figure
Sensitivity of the IceCube Detector to Astrophysical Sources of High Energy Muon Neutrinos
We present the results of a Monte-Carlo study of the sensitivity of the
planned IceCube detector to predicted fluxes of muon neutrinos at TeV to PeV
energies. A complete simulation of the detector and data analysis is used to
study the detector's capability to search for muon neutrinos from sources such
as active galaxies and gamma-ray bursts. We study the effective area and the
angular resolution of the detector as a function of muon energy and angle of
incidence. We present detailed calculations of the sensitivity of the detector
to both diffuse and pointlike neutrino emissions, including an assessment of
the sensitivity to neutrinos detected in coincidence with gamma-ray burst
observations. After three years of datataking, IceCube will have been able to
detect a point source flux of E^2*dN/dE = 7*10^-9 cm^-2s^-1GeV at a 5-sigma
significance, or, in the absence of a signal, place a 90% c.l. limit at a level
E^2*dN/dE = 2*10^-9 cm^-2s^-1GeV. A diffuse E-2 flux would be detectable at a
minimum strength of E^2*dN/dE = 1*10^-8 cm^-2s^-1sr^-1GeV. A gamma-ray burst
model following the formulation of Waxman and Bahcall would result in a 5-sigma
effect after the observation of 200 bursts in coincidence with satellite
observations of the gamma-rays.Comment: 33 pages, 13 figures, 6 table
Detection of Atmospheric Muon Neutrinos with the IceCube 9-String Detector
The IceCube neutrino detector is a cubic kilometer TeV to PeV neutrino
detector under construction at the geographic South Pole. The dominant
population of neutrinos detected in IceCube is due to meson decay in cosmic-ray
air showers. These atmospheric neutrinos are relatively well-understood and
serve as a calibration and verification tool for the new detector. In 2006, the
detector was approximately 10% completed, and we report on data acquired from
the detector in this configuration. We observe an atmospheric neutrino signal
consistent with expectations, demonstrating that the IceCube detector is
capable of identifying neutrino events. In the first 137.4 days of livetime,
234 neutrino candidates were selected with an expectation of 211 +/-
76.1(syst.) +/- 14.5(stat.) events from atmospheric neutrinos
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