2ign: Difference between revisions
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[[Image:2ign.gif|left|200px]] | [[Image:2ign.gif|left|200px]] | ||
'''Crystal structure of recombinant pyranose 2-oxidase H167A mutant''' | {{Structure | ||
|PDB= 2ign |SIZE=350|CAPTION= <scene name='initialview01'>2ign</scene>, resolution 1.65Å | |||
|SITE= | |||
|LIGAND= <scene name='pdbligand=FAD:FLAVIN-ADENINE+DINUCLEOTIDE'>FAD</scene> and <scene name='pdbligand=MES:2-(N-MORPHOLINO)-ETHANESULFONIC ACID'>MES</scene> | |||
|ACTIVITY= [http://en.wikipedia.org/wiki/Pyranose_oxidase Pyranose oxidase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=1.1.3.10 1.1.3.10] | |||
|GENE= p2o ([http://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?mode=Info&srchmode=5&id=230624 Trametes ochracea]) | |||
}} | |||
'''Crystal structure of recombinant pyranose 2-oxidase H167A mutant''' | |||
==Overview== | ==Overview== | ||
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==About this Structure== | ==About this Structure== | ||
2IGN is a [ | 2IGN is a [[Single protein]] structure of sequence from [http://en.wikipedia.org/wiki/Trametes_ochracea Trametes ochracea]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=2IGN OCA]. | ||
==Reference== | ==Reference== | ||
Structural basis for substrate binding and regioselective oxidation of monosaccharides at C3 by pyranose 2-oxidase., Kujawa M, Ebner H, Leitner C, Hallberg BM, Prongjit M, Sucharitakul J, Ludwig R, Rudsander U, Peterbauer C, Chaiyen P, Haltrich D, Divne C, J Biol Chem. 2006 Nov 17;281(46):35104-15. Epub 2006 Sep 19. PMID:[http:// | Structural basis for substrate binding and regioselective oxidation of monosaccharides at C3 by pyranose 2-oxidase., Kujawa M, Ebner H, Leitner C, Hallberg BM, Prongjit M, Sucharitakul J, Ludwig R, Rudsander U, Peterbauer C, Chaiyen P, Haltrich D, Divne C, J Biol Chem. 2006 Nov 17;281(46):35104-15. Epub 2006 Sep 19. PMID:[http://www.ncbi.nlm.nih.gov/pubmed/16984920 16984920] | ||
[[Category: Pyranose oxidase]] | [[Category: Pyranose oxidase]] | ||
[[Category: Single protein]] | [[Category: Single protein]] | ||
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[[Category: rossman fold]] | [[Category: rossman fold]] | ||
''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Mar 20 17:29:18 2008'' | ||
Revision as of 18:29, 20 March 2008
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| , resolution 1.65Å | |||||||
|---|---|---|---|---|---|---|---|
| Ligands: | and | ||||||
| Gene: | p2o (Trametes ochracea) | ||||||
| Activity: | Pyranose oxidase, with EC number 1.1.3.10 | ||||||
| Coordinates: | save as pdb, mmCIF, xml | ||||||
Crystal structure of recombinant pyranose 2-oxidase H167A mutant
OverviewOverview
Pyranose 2-oxidase (P2Ox) participates in fungal lignin degradation by producing the H2O2 needed for lignin-degrading peroxidases. The enzyme oxidizes cellulose- and hemicellulose-derived aldopyranoses at C2 preferentially, but also on C3, to the corresponding ketoaldoses. To investigate the structural determinants of catalysis, covalent flavinylation, substrate binding, and regioselectivity, wild-type and mutant P2Ox enzymes were produced and characterized biochemically and structurally. Removal of the histidyl-FAD linkage resulted in a catalytically competent enzyme containing tightly, but noncovalently bound FAD. This mutant (H167A) is characterized by a 5-fold lower kcat, and a 35-mV lower redox potential, although no significant structural changes were seen in its crystal structure. In previous structures of P2Ox, the substrate loop (residues 452-457) covering the active site has been either disordered or in a conformation incompatible with carbohydrate binding. We present here the crystal structure of H167A in complex with a slow substrate, 2-fluoro-2-deoxy-D-glucose. Based on the details of 2-fluoro-2-deoxy-D-glucose binding in position for oxidation at C3, we also outline a probable binding mode for D-glucose positioned for regioselective oxidation at C2. The tentative determinant for discriminating between the two binding modes is the position of the O6 hydroxyl group, which in the C2-oxidation mode can make favorable interactions with Asp452 in the substrate loop and, possibly, a nearby arginine residue (Arg472). We also substantiate our hypothesis with steady-state kinetics data for the alanine replacements of Asp452 and Arg472 as well as the double alanine 452/472 mutant.
About this StructureAbout this Structure
2IGN is a Single protein structure of sequence from Trametes ochracea. Full crystallographic information is available from OCA.
ReferenceReference
Structural basis for substrate binding and regioselective oxidation of monosaccharides at C3 by pyranose 2-oxidase., Kujawa M, Ebner H, Leitner C, Hallberg BM, Prongjit M, Sucharitakul J, Ludwig R, Rudsander U, Peterbauer C, Chaiyen P, Haltrich D, Divne C, J Biol Chem. 2006 Nov 17;281(46):35104-15. Epub 2006 Sep 19. PMID:16984920
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