393 research outputs found

    Multiplicity, Invariants and Tensor Product Decomposition of Tame Representations of U(\infty)

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    The structure of r-fold tensor products of irreducible tame representations of the inductive limit U(\infty) of unitary groups U(n) are are described, versions of contragredient representations and invariants are realized on Bargmann-Segal-Fock spaces.Comment: 48 pages, LaTeX file, to appear in J. Math. Phy

    Depsipeptide substrates for sortase-mediated N-terminal protein ligation

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    Technologies that allow the efficient chemical modification of proteins under mild conditions are widely sought after. Sortase-mediated peptide ligation provides a strategy for modifying the N or C terminus of proteins. This protocol describes the use of depsipeptide substrates (containing an ester linkage) with sortase A (SrtA) to completely modify proteins carrying a single N-terminal glycine residue under mild conditions in 4–6 h. The SrtA-mediated ligation reaction is reversible, so most labeling protocols that use this enzyme require a large excess of both substrate and sortase to produce high yields of ligation product. In contrast, switching to depsipeptide substrates effectively renders the reaction irreversible, allowing complete labeling of proteins with a small excess of substrate and catalytic quantities of sortase. Herein we describe the synthesis of depsipeptide substrates that contain an ester linkage between a threonine and glycolic acid residue and an N-terminal FITC fluorophore appended via a thiourea linkage. The synthesis of the depsipeptide substrate typically takes 2–3 d

    Anchoring of proteins to lactic acid bacteria

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    The anchoring of proteins to the cell surface of lactic acid bacteria (LAB) using genetic techniques is an exciting and emerging research area that holds great promise for a wide variety of biotechnological applications. This paper reviews five different types of anchoring domains that have been explored for their efficiency in attaching hybrid proteins to the cell membrane or cell wall of LAB. The most exploited anchoring regions are those with the LPXTG box that bind the proteins in a covalent way to the cell wall. In recent years, two new modes of cell wall protein anchoring have been studied and these may provide new approaches in surface display. The important progress that is being made with cell surface display of chimaeric proteins in the areas of vaccine development and enzyme- or whole-cell immobilisation is highlighted.

    Structural Differences between the Streptococcus agalactiae Housekeeping and Pilus-Specific Sortases: SrtA and SrtC1

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    The assembly of pili on the cell wall of Gram-positive bacteria requires transpeptidase enzymes called sortases. In Streptococcus agalactiae, the PI-1 pilus island of strain 2603V/R encodes two pilus-specific sortases (SrtC1 and SrtC2) and three pilins (GBS80, GBS52 and GBS104). Although either pilus-specific sortase is sufficient for the polymerization of the major pilin, GBS80, incorporation of the minor pilins GBS52 and GBS104 into the pilus structure requires SrtC1 and SrtC2, respectively. The S. agalactiae housekeeping sortase, SrtA, whose gene is present at a different location and does not catalyze pilus polymerization, was shown to be involved in cell wall anchoring of pilus polymers. To understand the structural basis of sortases involved in such diverse functions, we determined the crystal structures of S. agalactiae SrtC1 and SrtA. Both enzymes are made of an eight-stranded beta-barrel core with variations in their active site architecture. SrtA exhibits a catalytic triad arrangement similar to that in Streptococcus pyogenes SrtA but different from that in Staphylococcus aureus SrtA. In contrast, the SrtC1 enzyme contains an N-terminal helical domain and a ‘lid’ in its putative active site, which is similar to that seen in Streptococcus pneumoniae pilus-specific sortases, although with subtle differences in positioning and composition. To understand the effect of such differences on substrate recognition, we have also determined the crystal structure of a SrtC1 mutant, in which the conserved DP(W/F/Y) motif was replaced with the sorting signal motif of GBS80, IPNTG. By comparing the structures of WT wild type SrtA and SrtC1 and the ‘lid’ mutant of SrtC1, we propose that structural elements within the active site and the lid may be important for defining the role of specific sortase in pili biogenesis

    Nanostructured BiVO4 Photoanodes Fabricated by Vanadium-infused Interaction for Efficient Solar Water Splitting

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    Bismuth vanadate (BiVO4) has emerged as a highly prospective material for photoanodes in photoelectrochemical (PEC) water oxidation. However, current limitations with this material lie in the difficulties in producing stable and continuous BiVO4 layers with efficient carrier transfer kinetics, thereby impeding its widespread application in water splitting processes. This study introduces a new fabrication approach that yields continuous, monoclinic nanostructured BiVO4 films, paving the way for their use as photoanodes in efficient PEC water oxidation for hydrogen production under solar light conditions. The fabrication involves the intercalation of vanadium (V) ions into Bi2O3 films at 450oC. Upon interaction with V ions, the film undergoes a transformation from tetragonal Bi2O3 to monoclinic scheelite BiVO4. This synthesis method enables the fabrication of single monoclinic phase BiVO4 films with thicknesses up to 270 nm. The engineered monoclinic BiVO4 film, devoid of any pinholes that could cause carrier loss, exhibits a robust photocurrent of 1.0 mA/cm2 at 1.23 VRHE in a neutral electrolyte, without requiring additional modifications or doping. Moreover, we demonstrate that the incorporation of a cobalt phosphate (Co-Pi) co-catalyst into the BiVO4 photoanode significantly enhances the lifetime of photogenerated holes by a factor of nine, resulting in a further elevation of the photocurrent to 2.9 mA/cm2. This remarkable PEC enhancement can be attributed to the surface state passivation by the Co-Pi co-catalyst. Our fabrication approach opens up a new facile route for producing large-scale, highly efficient BiVO4 photoanodes for PEC water splitting technology

    Electronic structure of potassium-doped La@C_82 metallofullerene studied with photoelectron spectroscopy

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    The electronic structure of potassium-doped La@C_82 has been studied with synchrotron-radiation photoelectron spectroscopy. Ultraviolet photoemission measurements indicate evolution of the valence-band states of La@C_82 with increasing potassium content, but K_xLa@C_82 remains semiconducting for all x between 0 and 6, with a band gap of at least 0.4 eV, in contrast to K-doped C_60. The valence-band features, including those which arise from the interactions of C_82 cages with La and K, show oscillatory behavior of photoemission intensity with incident photon energy. The strength of the oscillations varies between the states, and we associate diminished oscillations with increased localization of electron density. Finally, although core and valence states of La@C_82 change considerably with potassium dosage, the encapsulated La atom is well shielded from the outside chemical environment and its valency remains close to 3+.journal articl

    Staphylococcus aureus forms spreading dendrites that have characteristics of active motility

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    Staphylococcus aureus is historically regarded as a non-motile organism. More recently it has been shown that S. aureus can passively move across agar surfaces in a process called spreading. We re-analysed spreading motility using a modified assay and fo- cused on observing the formation of dendrites: branching structures that emerge from the central colony. We discovered that S. aureus can spread across the surface of media in struc- tures that we term ‘comets’, which advance outwards and precede the formation of dendrites. We observed comets in a diverse selection of S. aureus isolates and they exhibit the following behaviours: (1) They consist of phenotypically distinct cores of cells that move forward and seed other S. aureus cells behind them forming a comet ‘tail’; (2) they move when other cells in the comet tail have stopped moving; (3) the comet core is held together by a matrix of slime; and (4) the comets etch trails in the agar as they move forwards. Comets are not con- sistent with spreading motility or other forms of passive motility. Comet behaviour does share many similarities with a form of active motility known as gliding. Our observations therefore suggest that S. aureus is actively motile under certain conditions

    Identification of sortase A (SrtA) substrates in Streptococcus uberis: evidence for an additional hexapeptide (LPXXXD) sorting motif

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    Sortase (a transamidase) has been shown to be responsible for the covalent attachment of proteins to the bacterial cell wall. Anchoring is effected on secreted proteins containing a specific cell wall motif toward their C-terminus; that for sortase A (SrtA) in Gram-positive bacteria often incorporates the sequence LPXTG. Such surface proteins are often characterized as virulence determinants and play important roles during the establishment and persistence of infection. Intramammary infection with Streptococcus uberis is a common cause of bovine mastitis, which impacts on animal health and welfare and the economics of milk production. Comparison of stringently produced cell wall fractions from S. uberis and an isogenic mutant strain lacking SrtA permitted identification of 9 proteins likely to be covalently anchored at the cell surface. Analysis of these sequences implied the presence of two anchoring motifs for S. uberis, the classical LPXTG motif and an additional LPXXXD motif
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