Abstract:
:Glycosynthases are engineered glycoside hydrolases (GHs) that catalyse the synthesis of glycoside from glycosyl-fluoride donors and suitable acceptors. We have determined five crystal structures of the glycosynthase mutants reducing-end xylose-releasing exo-oligoxylanase, an inverting GH, that exhibit various levels of glycosynthetic activities. At the active site of the Y198F mutant, the most efficient glycosynthase, a water molecule is observed at the same position as nucleophilic water (NW) in the parent enzyme, and the loss of the fixation of the direction of the lone pair of water molecules in the mutant drastically decreases hydrolytic activity. Water molecules were also observed at each active site of the general base mutant, but they were shifted 1.0-3.0 A from the NW in the wild type. Their positions exhibited a strong correlation with the strength of glycosynthase activity. Here, we propose that a structural prerequisite for the sufficient glycosynthase reaction is the presence of a water molecule at the NW position, and mutation at the NW holder provides a general strategy for inverting GHs. The idea on the position of a water molecule may also be applicable to the design of efficient glycosynthases from retaining GHs.
journal_name
J Biochemjournal_title
Journal of biochemistryauthors
Hidaka M,Fushinobu S,Honda Y,Wakagi T,Shoun H,Kitaoka Mdoi
10.1093/jb/mvp159subject
Has Abstractpub_date
2010-02-01 00:00:00pages
237-44issue
2eissn
0021-924Xissn
1756-2651pii
mvp159journal_volume
147pub_type
杂志文章abstract::The position of a single N'-formylkynurenine residue in ozone-inactivated hen egg-white lysozyme [EC 3.2.1.17] was determined by two methods. One involved identification of an amino-terminus of the C-peptide obtained by selective cleavage with hydrazine of the kynurenyl peptide linkage in oxidized lysozyme. The other ...
journal_title:Journal of biochemistry
pub_type: 杂志文章
doi:10.1093/oxfordjournals.jbchem.a131051
更新日期:1976-01-01 00:00:00
abstract::Fluorescence resonance energy transfer spectroscopy has been used to study the spatial relationships between probes attached to actin and troponin in the reconstituted skeletal muscle thin filament in the presence and absence of Ca2+ ions. Gln-41 and the nucleotide-binding site of actin were selectively labeled with t...
journal_title:Journal of biochemistry
pub_type: 杂志文章
doi:10.1093/oxfordjournals.jbchem.a021940
更新日期:1998-02-01 00:00:00
abstract::2,3-Bisphosphoglycerate mutase (BPGM) [EC 5.4.2.4] is a multifunctional enzyme that catalyzes both the synthesis and the degradation of 2,3-diphosphoglycerate (2,3-DPG) and contains three types of activities in that it functions as a 2,3-DPG synthetase, a phosphoglycerate mutase and a 2,3-DPG phosphatase. In humans, B...
journal_title:Journal of biochemistry
pub_type: 杂志文章
doi:10.1093/oxfordjournals.jbchem.a022243
更新日期:1998-12-01 00:00:00
abstract::The complete peptide map of purified recombinant human interleukin 5 (rhIL-5) was determined to verify its primary structure, glycosylation sites, and disulfide bonding structure. Each peptide fragment generated by Achromobacter protease I (API) digestion was purified and characterized by amino acid analysis and amino...
journal_title:Journal of biochemistry
pub_type: 杂志文章
doi:10.1093/oxfordjournals.jbchem.a123041
更新日期:1990-02-01 00:00:00
abstract::Aplyronine A is a macrolide isolated from Aplysia kurodai. By monitoring fluorescent intensity of pyrenyl-actin, it was found that aplyronine A inhibited both the velocity and the degree of actin polymerization. Aplyronine A also quickly depolymerized F-actin. The kinetics of depolymerization suggest that aplyronine A...
journal_title:Journal of biochemistry
pub_type: 杂志文章
doi:10.1093/oxfordjournals.jbchem.a021449
更新日期:1996-09-01 00:00:00
abstract::Polychlorinated biphenyls (PCBs) are well-known recalcitrant environmental pollutants. Although the metabolism of the PCBs has been intensively studied, very little is known about their mechanism of toxicity in living organisms or how they are degraded. We have examined the effects of PCBs on two different yeast strai...
journal_title:Journal of biochemistry
pub_type: 杂志文章
doi:10.1093/jb/mvg121
更新日期:2003-07-01 00:00:00
abstract::The structure and expression of the pentraxins, serum amyloid P component (SAP), and C-reactive protein (CRP), have been investigated in the guinea pig. Northern blot analysis of hepatic RNA from animals in which acute inflammation had been induced by intraperitoneal injection of thioglycollate established that neithe...
journal_title:Journal of biochemistry
pub_type: 杂志文章
doi:10.1093/oxfordjournals.jbchem.a124039
更新日期:1993-03-01 00:00:00
abstract::Aberrant DNA methylation is a common form of epigenetic alterations and it has been proved to be closely related to many cancers, while its role in epidermal growth factor receptor (EGFR)-mutated non-small cell lung cancer (NSCLC) is not clear. This study focuses on the role of DNA methyltransferase 1 (DNMT1) in EGFR-...
journal_title:Journal of biochemistry
pub_type: 杂志文章
doi:10.1093/jb/mvaa034
更新日期:2020-08-01 00:00:00
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journal_title:Journal of biochemistry
pub_type: 杂志文章
doi:10.1093/oxfordjournals.jbchem.a022587
更新日期:2000-01-01 00:00:00
abstract::Three isozymic forms, alpha, beta, and gamma, of Drosophila melanogaster aldolase are produced from a single gene by alternative usage of the triple exons 4 (4 alpha, 4 beta, and 4 gamma) [Shaw-Lee et al. (1992) J. Biol. Chem. 267, 3959-3967; Kim et al. (1992) Mol. Cell. Biol. 12, 773-783; Kai et al. (1992) J. Biochem...
journal_title:Journal of biochemistry
pub_type: 杂志文章
doi:10.1093/oxfordjournals.jbchem.a124876
更新日期:1995-07-01 00:00:00
abstract::Mitochondria play many critical roles in cells, not only by supplying energy, but also by supplying metabolites, buffering Ca2+ levels and regulating apoptosis. During oocyte maturation and subsequent embryo development, mitochondria change their morphology by membrane fusion and fission, and coordinately undergo mult...
journal_title:Journal of biochemistry
pub_type: 杂志文章,评审
doi:10.1093/jb/mvz093
更新日期:2020-03-01 00:00:00
abstract::Formate dehydrogenase (EC 1.2.1.2) prepared from peas (Pisum sativum) was a two-subunit enzyme. The enzyme accelerated the formation of an NAD+-cyanide compound having an adsorption band at 330 nm. The enzyme was able to bind one NAD+ molecule per each subunit but only 1 mole of NAD+-cyanide compound was formed per ...
journal_title:Journal of biochemistry
pub_type: 杂志文章
doi:10.1093/oxfordjournals.jbchem.a130792
更新日期:1975-04-01 00:00:00
abstract::X-ray diffraction photographs of a chicken gizzard G-actin.DNase I complex crystal have been recorded using the synchrotron radiation beam emitted by the Synchrotron Radiation Source at Daresbury and the Photon Factory at Tsukuba. The resolution limit was extended to 2.4 A and the exposure time was reduced approximate...
journal_title:Journal of biochemistry
pub_type: 杂志文章
doi:10.1093/oxfordjournals.jbchem.a134682
更新日期:1984-03-01 00:00:00
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journal_title:Journal of biochemistry
pub_type: 杂志文章
doi:10.1093/oxfordjournals.jbchem.a002896
更新日期:2001-04-01 00:00:00
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journal_title:Journal of biochemistry
pub_type: 杂志文章
doi:10.1093/oxfordjournals.jbchem.a021254
更新日期:1996-03-01 00:00:00
abstract::Pollen proteins from several grass species have been identified and characterized as causative allergens in grass pollinosis. In contrast, allergenic potential of pollen proteins from rice, which belongs to the same Poaceae family, has not well been investigated, despite that a few clinical cases have been reported on...
journal_title:Journal of biochemistry
pub_type: 临床试验,杂志文章
doi:10.1093/jb/mvt044
更新日期:2013-08-01 00:00:00
abstract::This study presents the structural, kinetic and thermodynamic characterizations of previously unknown submicellar concentrations of SDS-induced molten globule (MGSDS) state of a highly negatively charged base-denatured ferricytochrome c (UB-state) at pH ∼12.8 (±0.2). The far-UV CD, near-UV CD, ANS-fluorescence data of...
journal_title:Journal of biochemistry
pub_type: 杂志文章
doi:10.1093/jb/mvy087
更新日期:2019-02-01 00:00:00
abstract::We purified dipeptidyl peptidase II (DPP II) to homogeneity from rat kidney and determined its physicochemical properties, including its molecular weight, substrate specificity, and partial amino acid sequence. Furthermore, we screened a rat kidney cDNA library, isolated the DPP II cDNA and determined its structure. T...
journal_title:Journal of biochemistry
pub_type: 杂志文章
doi:10.1093/oxfordjournals.jbchem.a002855
更新日期:2001-02-01 00:00:00
abstract::1. Glyceraldehyde-3-phosphate dehydrogenases [EC 1.2.1.12] from rabbit, pig, lobster, yeast, E. coli, and B. stearothermophilus have been subjected to hybridization in 3M NaCL. 2. Suitable mixtures of electrophoretically distinct glyceraldehyde-3-phosphate dehydrogenases were found to give five-membered by electrophor...
journal_title:Journal of biochemistry
pub_type: 杂志文章
doi:10.1093/oxfordjournals.jbchem.a130760
更新日期:1975-03-01 00:00:00
abstract::Vasoactive intestinal peptide (VIP), a neuropeptide coupled with adenylate cyclase, was found to induce neurite extension of PC12h cells. Neurites appeared within 1 h after addition of VIP and extended for at least 24 h. The half-maximal concentration for the effect of VIP was 50 nM. In addition to the morphological c...
journal_title:Journal of biochemistry
pub_type: 杂志文章
doi:10.1093/oxfordjournals.jbchem.a124333
更新日期:1994-02-01 00:00:00
abstract::We tested the capsid targeted viral inactivation method as an anti-HBV strategy. HepG2 cells were cotransfected with HBV expression plasmid and the plasmid encoding fusion protein of either Core-A3C or Core-humanized renilla GFP (hrGFP). Core-A3C had substantial effect on HBV DNA levels. In the HepG2 cells expressing ...
journal_title:Journal of biochemistry
pub_type: 杂志文章
doi:10.1093/jb/mvr086
更新日期:2011-10-01 00:00:00
abstract::We have studied the protein composition of the pectoralis superficialis muscle of genetically dystrophic (New Hampshire line 413) and normal control (line 412) chickens by one- and two-dimensional gel electrophoresis. A protein, referred to hereafter as the 30 kDa abnormal protein, was specifically detected in the aff...
journal_title:Journal of biochemistry
pub_type: 杂志文章
doi:10.1093/oxfordjournals.jbchem.a123010
更新日期:1990-01-01 00:00:00
abstract::Guinea pig alveolar macrophages (A-M0) were studied with respect ot the phagocytic metabolism: the respiratory burst and its KCN sensitivity, the ability to generate H2O2, and myeloperoxidase (MPO) activity. Only fifty percent of the O2 uptake by A-M0 samples during phagocytosis was inhibited by the addition of 1 mK K...
journal_title:Journal of biochemistry
pub_type: 杂志文章
doi:10.1093/oxfordjournals.jbchem.a133070
更新日期:1980-10-01 00:00:00
abstract::Treatment of Sendai virus with p-azidophenyl-6-chloro-6-deoxy-beta-D-glucopyranoside (APG) caused chemical modification of the viral envelope under UV irradiation, which did not affect the hemagglutinin activity of the virus but inhibited the hemolytic activity. Also, the transfer of phospholipid from the viral envelo...
journal_title:Journal of biochemistry
pub_type: 杂志文章
doi:10.1093/oxfordjournals.jbchem.a133088
更新日期:1980-10-01 00:00:00
abstract::In the rat, DNA carrying a cluster of the genes for tRNAAsp, tRNAGly, and tRNAGlu, aligned in that order, is repeated about 10 times. Seven DNA clones corresponding to the independent repeating units were isolated from a rat gene library. Nucleotide sequence analysis of these clones revealed the presence of a fourth t...
journal_title:Journal of biochemistry
pub_type: 杂志文章
doi:10.1093/oxfordjournals.jbchem.a135230
更新日期:1985-06-01 00:00:00
abstract::GM2 containing NeuGc was found to be a major ganglioside in the liver of most inbred strains of mouse we have examined so far. They were DBA/2, C3H/He, C57BL/6, BALB/c, and CBA mice. However, WHT/Ht mouse does not contain a detectable amount of GM2 (NeuGc) but has GM3 containing NeuGc as the major ganglioside in the l...
journal_title:Journal of biochemistry
pub_type: 杂志文章
doi:10.1093/jb/93.3.895
更新日期:1983-03-01 00:00:00
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journal_title:Journal of biochemistry
pub_type: 杂志文章
doi:10.1093/jb/mvu042
更新日期:2014-12-01 00:00:00
abstract::The amino acid sequences of ferredoxins I and II from a blue-green alga, Nostoc strain MAC were determined. This alga is able to grow autotrophically in the light or heterotrophically in the dark. Analyses of tryptic peptides of Cm-proteins by conventional methods including solid-phase Edman degradation gave the compl...
journal_title:Journal of biochemistry
pub_type: 杂志文章
doi:10.1093/oxfordjournals.jbchem.a134058
更新日期:1982-11-01 00:00:00
abstract::Sugars such as glycolaldehyde, glyceraldehyde, erythrose, ribose, and glucose were heated with ammonia in a modified sea medium. Amino acids such as glycine, alanine, serine, threonine, aspartic acid, and glutamic acid were obtained from the reaction mixture. This provides the basis for a model of the genesis of amino...
journal_title:Journal of biochemistry
pub_type: 杂志文章
doi:10.1093/oxfordjournals.jbchem.a132748
更新日期:1980-01-01 00:00:00
abstract::The position in the acyl phosphate linkage of the phosphorylated intermediate of (Na+, K+)-ATPase that is cleaved by N-methylhydroxylamine was compared with that of the model compound acetylphosphate. The products of the cleavage of the phosphoenzyme by methylhydroxylamine were the active enzyme and a N-P compound, no...
journal_title:Journal of biochemistry
pub_type: 杂志文章
doi:10.1093/oxfordjournals.jbchem.a021805
更新日期:1997-09-01 00:00:00