220 research outputs found
Bulk angular momentum and Hall viscosity in chiral superconductors
We establish the Berry-phase formulas for the angular momentum (AM) and the
Hall viscosity (HV) to investigate chiral superconductors (SCs) in two and
three dimensions. The AM is defined by the temporal integral of the
anti-symmetric momentum current induced by an adiabatic deformation, while the
HV is defined by the symmetric momentum current induced by the symmetric
torsional electric field. Without suffering from the system size or geometry,
we obtain the macroscopic AM at zero temperature in
full-gap chiral SCs, where is the magnetic quantum number and is the
total number of electrons. We also find that the HV is equal to half the AM at
zero temperature not only in full-gap chiral SCs as is well-known but also in
nodal ones, but its behavior at finite temperature is different in the two
cases.Comment: 10 pages, 4 figures, 1 tabl
Replacement of the Catalytic Nucleophile Aspartyl Residue of Dextran Glucosidase by Cysteine Sulfinate Enhances Transglycosylation Activity
Dextran glucosidase from Streptococcus mutans (SmDG) catalyzes the hydrolysis of an α-1,6-glucosidic linkage at the nonreducing end of isomaltooligosaccharides and dextran. This enzyme has an Asp-194 catalytic nucleophile and two catalytically unrelated Cys residues, Cys-129 and Cys-532. Cys-free SmDG was constructed by replacement with Ser (C129S/C532S (2CS), the activity of which was the same as that of the wild type, SmDG). The nucleophile mutant of 2CS was generated by substitution of Asp-194 with Cys (D194C-2CS). The hydrolytic activity of D194C-2CS was 8.1 × 10⁻⁴ % of 2CS. KI-associated oxidation of D194C-2CS increased the activity up to 0.27% of 2CS, which was 330 times higher than D194C-2CS. Peptide-mapping mass analysis of the oxidized D194C-2CS (Ox-D194C-2CS) revealed that Cys-194 was converted into cysteine sulfinate. Ox-D194C-2CS and 2CS shared the same properties (optimum pH, pI, and substrate specificity), whereas Ox-D194C-2CS had much higher transglucosylation activity than 2CS. This is the first study indicating that a more acidic nucleophile (-SOO−) enhances transglycosylation. The introduction of cysteine sulfinate as a catalytic nucleophile could be a novel approach to enhance transglycosylation
Characterization of some enzymatic properties of recombinant α-glucosidase III from the Thai honeybee, Apis cerana indica Fabricus
Recombinant α-glucosidase III (rHBGase III) from Apis cerana indica Fabricus (rAciHBGase III) was expressed in the yeast Pichia pastoris GS115, enriched and characterized. The full length cDNA of AciHbgase III (ƒî1.8 kb) was amplified by RT-PCR, cloned into the pPICZαA expression vector and used to transform P. pastoris GS115. The maximum secreted expression level of rAciHBGase III [as an N terminal (His)6 tagged chimera] was found 144 h after induction by 1% (v/v) methanol. Enrichment of the enzyme using histrap affinity purification revealed a single active glucosidase band with a molecular mass of ~68 kDa. The optimal pH and temperature for glucosidase activity of the enriched rAciHBGase III were pH 5.0 and 37¢XC, respectively, whilst the enzyme showed a good pH stability between pH 5.0 to 7.5, but not below pH 5.0, and a poor thermotolerance with < 10% and 0% residual activity at 40 and >50¢XC, respectively. The rAciHBGase showed a relatively high substrate specificity for maltose (Km of 4.5 mM) and p-nitrophenyl α-D-glucoside (Km of 4.4 mM) compared to other reported HBGase enzymes.Key words: α-Glucosidase, Apis cerana indica, expression, kinetics, recombinant enzyme
Catalytic role of the calcium ion in GH97 inverting glycoside hydrolase
AbstractThe role of calcium ion in the active site of the inverting glycoside hydrolase family 97 enzyme, BtGH97a, was investigated through structural and kinetic studies. The calcium ion was likely directly involved in the catalytic reaction. The pH dependence of kcat/Km values in the presence or absence of calcium ion indicated that the calcium ion lowered the pKa of the base catalyst. The significant decreases in kcat/Km for hydrolysis of substrates with basic leaving groups in the absence of calcium ion confirmed that the calcium ion facilitated the leaving group departure
One-step generation of multiple transgenic mouse lines using an improved Pronuclear Injection-based Targeted Transgenesis (i-PITT)
Ohtsuka, M., Miura, H., Mochida, K. et al. One-step generation of multiple transgenic mouse lines using an improved Pronuclear Injection-based Targeted Transgenesis (i-PITT). BMC Genomics 16, 274 (2015). https://doi.org/10.1186/s12864-015-1432-
An Initial Step of GAS-Containing Autophagosome-Like Vacuoles Formation Requires Rab7
Group A streptococcus (GAS; Streptococcus pyogenes) is a common pathogen that invades non-phagocytic human cells via endocytosis. Once taken up by cells, it escapes from the endocytic pathway to the cytoplasm, but here it is contained within a membrane-bound structure termed GAS-containing autophagosome-like vacuoles (GcAVs). The autophagosome marker GFP-LC3 associates with GcAVs, and other components of the autophagosomal pathway are involved in GcAV formation. However, the mechanistic relationship between GcAV and canonical autophagy is largely unknown. Here, we morphologically analyzed GcAV formation in detail. Initially, a small, GFP-LC3-positive GcAV sequesters each streptococcal chain, and these then coalesce into a single, large GcAV. Expression of a dominant-negative form of Rab7 or RNAi-mediated knockdown of Rab7 prevented the initial formation of small GcAV structures. Our results demonstrate that mechanisms of GcAV formation includes not only the common machinery of autophagy, but also Rab7 as an additional component, which is dispensable in canonical autophagosome formation
Randomized Comparison Between Everolimus-Eluting Bioresorbable Scaffold and Metallic Stent: Multimodality Imaging Through 3 Years
Objectives: The aim of this study was to investigate the vascular responses and fates of the scaffold after bioresorbable vascular scaffold (BVS) implantation using multimodality imaging. Background: Serial comprehen
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