2gc3: Difference between revisions

Revision as of 18:30, 21 February 2008

The crystal structure of phosphoglucose isomerase from Pyrococcus furiosus in complex with mannose 6-phosphate and zinc

OverviewOverview

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.

About this StructureAbout this Structure

2GC3 is a Single protein structure of sequence from Pyrococcus furiosus with and as ligands. Active as Glucose-6-phosphate isomerase, with EC number 5.3.1.9 Full crystallographic information is available from OCA.

ReferenceReference

Evidence supporting a cis-enediol-based mechanism for Pyrococcus furiosus phosphoglucose isomerase., Berrisford JM, Hounslow AM, Akerboom J, Hagen WR, Brouns SJ, van der Oost J, Murray IA, Michael Blackburn G, Waltho JP, Rice DW, Baker PJ, J Mol Biol. 2006 May 19;358(5):1353-66. Epub 2006 Mar 24. PMID:16580686

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-[[Image:2gc3.gif|left|200px]]<br /><applet load="2gc3" size="450" color="white" frame="true" align="right" spinBox="true"
+[[Image:2gc3.gif|left|200px]]<br /><applet load="2gc3" size="350" color="white" frame="true" align="right" spinBox="true"
 caption="2gc3, resolution 2.100&Aring;" />
 '''The crystal structure of phosphoglucose isomerase from Pyrococcus furiosus in complex with mannose 6-phosphate and zinc'''<br />
 ==Overview==
-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.
+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.
 ==About this Structure==
-GC3 is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Pyrococcus_furiosus Pyrococcus furiosus] with ZN and M6P as [http://en.wikipedia.org/wiki/ligands ligands]. Active as [http://en.wikipedia.org/wiki/Glucose-6-phosphate_isomerase Glucose-6-phosphate isomerase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=5.3.1.9 5.3.1.9] Full crystallographic information is available from [http://ispc.weizmann.ac.il/oca-bin/ocashort?id=2GC3 OCA].
+GC3 is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Pyrococcus_furiosus Pyrococcus furiosus] with <scene name='pdbligand=ZN:'>ZN</scene> and <scene name='pdbligand=M6P:'>M6P</scene> as [http://en.wikipedia.org/wiki/ligands ligands]. Active as [http://en.wikipedia.org/wiki/Glucose-6-phosphate_isomerase Glucose-6-phosphate isomerase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=5.3.1.9 5.3.1.9] Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=2GC3 OCA].
 ==Reference==
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 [[Category: Pyrococcus furiosus]]
 [[Category: Single protein]]
-[[Category: Baker, P.J.]]
+[[Category: Baker, P J.]]
-[[Category: Berrisford, J.M.]]
+[[Category: Berrisford, J M.]]
-[[Category: Rice, D.W.]]
+[[Category: Rice, D W.]]
 [[Category: M6P]]
 [[Category: ZN]]
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 [[Category: phosphoglucose isomerase]]
-''Page seeded by [http://ispc.weizmann.ac.il/oca OCA ] on Wed Nov 21 11:06:33 2007''
+''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 17:30:15 2008''