Staff Publications

Staff Publications

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    'Staff publications' is the digital repository of Wageningen University & Research

    'Staff publications' contains references to publications authored by Wageningen University staff from 1976 onward.

    Publications authored by the staff of the Research Institutes are available from 1995 onwards.

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Record number 346219
Title Evidence supporting a cis-enediol-based mechanism for Pyrococcus furiosus phosphoglucose isomerase
Author(s) Berrisford, J.M.; Hounslow, A.M.; Akerboom, A.P.; Hagen, W.R.; Brouns, S.J.J.; Oost, J. van der; Murray, I.A.; Blackburn, G.M.; Waltho, J.P.; Rice, D.W.; Baker, P.J.
Source Journal of Molecular Biology 358 (2006)5. - ISSN 0022-2836 - p. 1353 - 1366.
DOI https://doi.org/10.1016/j.jmb.2006.03.015
Department(s) Microbiological Laboratory
VLAG
Publication type Refereed Article in a scientific journal
Publication year 2006
Keyword(s) crystal-structure - substrate-binding - glucose-6-phosphate isomerase - phosphomannose isomerase - molecular-replacement - enzyme - inhibitor - complexes - catalysis - crystallization
Abstract The enzymatic aldose ketose isomerisation of glucose and fructose sugars involves the transfer of a hydrogen between their C1 and C2 carbon atoms and, in principle, can proceed through either a direct hydride shift or via a cis-enediol intermediate. Pyrococcus furiosus phosphoglucose isomerase (PfPGI), an archaeal metalloenzyme, which catalyses the interconversion of glucose 6-phosphate and fructose 6-phosphate, has been suggested to operate via a hydride shift mechanism. In contrast, the structurally distinct PGIs of eukaryotic or bacterial origin are thought to catalyse isomerisation via a cis-enediol intermediate. We have shown by NMR that hydrogen exchange between substrate and solvent occurs during the reaction catalysed by PfPGI eliminating the possibility of a hydride-shift-based mechanism. In addition, kinetic measurements on this enzyme have shown that 5-phospho-d-arabinonohydroxamate, a stable analogue of the putative cis-enediol intermediate, is the most potent inhibitor of the enzyme yet discovered. Furthermore, determination and analysis of crystal structures of PfPGI with bound zinc and the substrate F6P, and with a number of competitive inhibitors, and EPR analysis of the coordination of the metal ion within PfPGI, have suggested that a cis-enediol intermediate-based mechanism is used by PfPGI with Glu97 acting as the catalytic base responsible for isomerisation.
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