Structural studies of the retaining galactosyltransferase LGTC from Neisseria meningitidis

Teeri, Tuula T.; Svensson, Birte; Gilbert, Harry J.; Feizi, Ten; Persson, Karina; Ly, Hoa D.; Dickelmann, M.; Wakarchuk, Warren W.; Withers, Stephen G.; Strynadka, N.C.J.

doi:10.1039/9781847550323-00042

Cited by 74 publications

(174 citation statements)

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“…The notion that α-retaining glycosyl transfer catalysed by phosphorylases and nucleotide-dependent GTs could occur generally without the requirement for a covalent β-glycosyl-enzyme intermediate [30] has gained considerable support from the recent X-ray structures of maltodextrin phosphorylase from Escherichia coli [31] and galactosyltransferase LtgC from Neisseria meningitidis [27] in complexes with donor and acceptor analogues. Inspection of the active sites of both enzymes revealed clearly that no carboxylate group is placed into position as a candidate catalytic nucleophile.…”

Section: Kinetic Mechanism Of Trehalose Phosphorylasementioning

confidence: 99%

“…Bivalent metal ions such as Mg# + or Mn# + have been shown to be co-ordinated to two oxygen atoms from each of the two phosphates in nucleotide diphosphates, and interact with an acidic amino acid, usually an aspartate from a widely conserved Asp-Xaa-Asp motif [27,[36][37][38]. Water molecules complete the co-ordination sphere of the metal ion.…”

Section: Role Of Enzyme-bound Mg 2 +mentioning

confidence: 99%

“…Inspection of the active sites of both enzymes revealed clearly that no carboxylate group is placed into position as a candidate catalytic nucleophile. Since evidence for a doubledisplacement mechanism has remained elusive for maltodextrin phosphorylase and galactosyltransferase, alternative mechanistic scenarios have been proposed [11,12,15,27,30,31]. One of these is the so-called internal return (S N i) mechanism of glycosyl transfer that assumes approach of the nucleophile towards the reactive carbon from the same side from where the leaving group departs [11,32].…”

Section: Kinetic Mechanism Of Trehalose Phosphorylasementioning

confidence: 99%

“…Mg# + (or Mn# + ) may play an important role in the catalytic mechanism of glycosyl transfer catalysed by retaining [27,30] and inverting nucleotide sugar-dependent transferases (reviewed in [36] ; see also [37,38]), which are related to trehalose phosphorylase through family relationships (GT family 4) or general mechanistic similarities (see [11] for a detailed discussion of mechanisms). Bivalent metal ions such as Mg# + or Mn# + have been shown to be co-ordinated to two oxygen atoms from each of the two phosphates in nucleotide diphosphates, and interact with an acidic amino acid, usually an aspartate from a widely conserved Asp-Xaa-Asp motif [27,[36][37][38].…”

Section: Role Of Enzyme-bound Mg 2 +mentioning

confidence: 99%

“…A few other α-retaining glucosyl transferases are known to utilize sequential kinetic mechanisms that are characterized by the strict requirement for a ternary enzyme-substrate complex : sucrose synthetase (GT family 4) [23,24], glycogen synthetase (GT family 5) [25,26], galactosyltransferase LgtC (GT family 8) [27] and glycogen phosphorylase (GT family 35) [28]. It has been speculated that ternary-complex kinetics could reflect an enzymic mechanism for minimizing the likelihood of the reaction of a glucosyl-enzyme intermediate (or another reactive intermediate) with water and thus catalysing glucosyl transfer with only scant hydrolysis.…”

Section: Kinetic Mechanism Of Trehalose Phosphorylasementioning

confidence: 99%

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Fungal trehalose phosphorylase: kinetic mechanism, pH-dependence of the reaction and some structural properties of the enzyme from Schizophyllum commune

et al. 2001

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Initial-velocity measurements for the phospholysis and synthesis of α,α-trehalose catalysed by trehalose phosphorylase from Schizophyllum commune and product and dead-end inhibitor studies show that this enzyme has an ordered Bi Bi kinetic mechanism, in which phosphate binds before α,α-trehalose, and α--glucose is released before α--glucose 1-phosphate. The freeenergy profile for the enzymic reaction at physiological reactant concentrations displays its largest barriers for steps involved in reverse glucosyl transfer to -glucose, and reveals the direction of phospholysis to be favoured thermodynamically. The pH dependence of kinetic parameters for all substrates and the dissociation constant of -glucal, a competitive dead-end inhibitor against -glucose (K i l 0.3 mM at pH 6.6 and 30 mC), were determined. Maximum velocities and catalytic efficiencies for the forward and reverse reactions decrease at high and low pH, giving apparent pK values of 7.2-7.8 and 5.5-6.0 for two groups whose correct protonation state is required for catalysis. The pH dependences of k cat \K are interpreted in terms of

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Section: Kinetic Mechanism Of Trehalose Phosphorylasementioning

confidence: 99%

Section: Role Of Enzyme-bound Mg 2 +mentioning

confidence: 99%

Section: Kinetic Mechanism Of Trehalose Phosphorylasementioning

confidence: 99%

Section: Role Of Enzyme-bound Mg 2 +mentioning

confidence: 99%

Section: Kinetic Mechanism Of Trehalose Phosphorylasementioning

confidence: 99%

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Fungal trehalose phosphorylase: kinetic mechanism, pH-dependence of the reaction and some structural properties of the enzyme from Schizophyllum commune

et al. 2001

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Chemoenzymatic synthesis of 6‐phospho‐cyclophellitol as a novel probe of 6‐phospho‐β‐glucosidases

et al. 2016

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Edited by Stuart FergusonCovalent, mechanism-based inhibitors of glycosidases are valuable probe molecules for visualizing enzyme activities in complex systems. We, here, describe the chemoenzymatic synthesis of 6-phospho-cyclophellitol and evaluate its behaviour as a mechanism-based inactivator of the Streptococcus pyogenes 6-phospho-b-glucosidase from CAZy family GH1. We further present the three-dimensional structure of the inactivated enzyme, which reveals the constellation of active site residues responsible for the enzyme's specificity and confirms the covalent nature of the inactivation.

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Glycosyltransferases of the GT8 Family

Yin¹,

Mohnen²,

Gelineo-Albersheim³

et al. 2010

Annual Plant Reviews

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The higher plant genomes sequenced to date include numerous genes encoding proteins classifi ed as belonging to CAZy family GT8. The large number and diversity of GT8 proteins in higher plants, which currently constitute more than 65% of the identifi ed eukaryotic GT8 genes, highlight the importance of these proteins in plants. Here we summarize a detailed phylogenetic study of GT8 proteins from three monocot and four dicot plant genomes that clearly divides higher plant GT8 proteins into two distantly related sets of clades, many of which are further divided into statistically well -supported subclades. One set, the GAUT1 ( GA lact U ronosyl T ransferase1) -related family, includes the GAUT and GA U T -L ike (GATL) proteins, comprising one proven galacturonsyltransferase and multiple additional members strongly implicated in the synthesis of pectins and xylan, two major types of polysaccharides present in plant cell walls. The second set, which includes P lant G lycogenin -like S tarch I nitiation P rotein s (PGSIPs) and G alactin ol S ynthase s (GolSs), appears not to be directly involved in plant cell wall synthesis. The PGSIPs have been suggested to play a role in priming starch biosynthesis, while the GolSs are key enzymes in the synthesis of

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Structural studies of the retaining galactosyltransferase LGTC from Neisseria meningitidis

Cited by 74 publications

References 0 publications

Fungal trehalose phosphorylase: kinetic mechanism, pH-dependence of the reaction and some structural properties of the enzyme from Schizophyllum commune

Fungal trehalose phosphorylase: kinetic mechanism, pH-dependence of the reaction and some structural properties of the enzyme from Schizophyllum commune

Chemoenzymatic synthesis of 6‐phospho‐cyclophellitol as a novel probe of 6‐phospho‐β‐glucosidases

Glycosyltransferases of the GT8 Family

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