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| - | [[Image:2ign.gif|left|200px]] | |
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| - | {{Structure
| + | ==Crystal structure of recombinant pyranose 2-oxidase H167A mutant== |
| - | |PDB= 2ign |SIZE=350|CAPTION= <scene name='initialview01'>2ign</scene>, resolution 1.65Å
| + | <StructureSection load='2ign' size='340' side='right'caption='[[2ign]], [[Resolution|resolution]] 1.65Å' scene=''> |
| - | |SITE=
| + | == Structural highlights == |
| - | |LIGAND= <scene name='pdbligand=FAD:FLAVIN-ADENINE+DINUCLEOTIDE'>FAD</scene> and <scene name='pdbligand=MES:2-(N-MORPHOLINO)-ETHANESULFONIC ACID'>MES</scene> | + | <table><tr><td colspan='2'>[[2ign]] is a 8 chain structure with sequence from [https://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]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=2IGN FirstGlance]. <br> |
| - | |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]
| + | </td></tr><tr id='method'><td class="sblockLbl"><b>[[Empirical_models|Method:]]</b></td><td class="sblockDat" id="methodDat">X-ray diffraction, [[Resolution|Resolution]] 1.65Å</td></tr> |
| - | |GENE= p2o ([http://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?mode=Info&srchmode=5&id=230624 Trametes ochracea]) | + | <tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=FAD:FLAVIN-ADENINE+DINUCLEOTIDE'>FAD</scene>, <scene name='pdbligand=MES:2-(N-MORPHOLINO)-ETHANESULFONIC+ACID'>MES</scene></td></tr> |
| - | }}
| + | <tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[https://proteopedia.org/fgij/fg.htm?mol=2ign FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=2ign OCA], [https://pdbe.org/2ign PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=2ign RCSB], [https://www.ebi.ac.uk/pdbsum/2ign PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=2ign ProSAT]</span></td></tr> |
| | + | </table> |
| | + | == Function == |
| | + | [https://www.uniprot.org/uniprot/Q7ZA32_TRAOC Q7ZA32_TRAOC] |
| | + | == Evolutionary Conservation == |
| | + | [[Image:Consurf_key_small.gif|200px|right]] |
| | + | Check<jmol> |
| | + | <jmolCheckbox> |
| | + | <scriptWhenChecked>; select protein; define ~consurf_to_do selected; consurf_initial_scene = true; script "/wiki/ConSurf/ig/2ign_consurf.spt"</scriptWhenChecked> |
| | + | <scriptWhenUnchecked>script /wiki/extensions/Proteopedia/spt/initialview01.spt</scriptWhenUnchecked> |
| | + | <text>to colour the structure by Evolutionary Conservation</text> |
| | + | </jmolCheckbox> |
| | + | </jmol>, as determined by [http://consurfdb.tau.ac.il/ ConSurfDB]. You may read the [[Conservation%2C_Evolutionary|explanation]] of the method and the full data available from [http://bental.tau.ac.il/new_ConSurfDB/main_output.php?pdb_ID=2ign ConSurf]. |
| | + | <div style="clear:both"></div> |
| | | | |
| - | '''Crystal structure of recombinant pyranose 2-oxidase H167A mutant'''
| + | ==See Also== |
| - | | + | *[[Pyranose oxidase|Pyranose oxidase]] |
| - | | + | __TOC__ |
| - | ==Overview== | + | </StructureSection> |
| - | 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. | + | [[Category: Large Structures]] |
| - | | + | |
| - | ==About this Structure==
| + | |
| - | 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].
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| - | | + | |
| - | ==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://www.ncbi.nlm.nih.gov/pubmed/16984920 16984920]
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| - | [[Category: Pyranose oxidase]]
| + | |
| - | [[Category: Single protein]] | + | |
| | [[Category: Trametes ochracea]] | | [[Category: Trametes ochracea]] |
| - | [[Category: Divne, C.]] | + | [[Category: Divne C]] |
| - | [[Category: FAD]]
| + | |
| - | [[Category: MES]]
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| - | [[Category: 8-alpha-(n3) histidyl flavinylation]]
| + | |
| - | [[Category: gmc oxidoreductase]]
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| - | [[Category: h167a mutant]]
| + | |
| - | [[Category: homotetramer]]
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| - | [[Category: oxidoreductase]]
| + | |
| - | [[Category: phbh fold]]
| + | |
| - | [[Category: rossman fold]]
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| - | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Mar 20 17:29:18 2008''
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