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| [[Image:1pbd.gif|left|200px]]<br /><applet load="1pbd" size="450" color="white" frame="true" align="right" spinBox="true"
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| caption="1pbd, resolution 2.30Å" />
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| '''CRYSTAL STRUCTURES OF WILD-TYPE P-HYDROXYBENZOATE HYDROXYLASE COMPLEXED WITH 4-AMINOBENZOATE, 2,4-DIHYDROXYBENZOATE AND 2-HYDROXY-4-AMINOBENZOATE AND OF THE TRY222ALA MUTANT, COMPLEXED WITH 2-HYDROXY-4-AMINOBENZOATE. EVIDENCE FOR A PROTON CHANNEL AND A NEW BINDING MODE OF THE FLAVIN RING'''<br />
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| ==Overview== | | ==CRYSTAL STRUCTURES OF WILD-TYPE P-HYDROXYBENZOATE HYDROXYLASE COMPLEXED WITH 4-AMINOBENZOATE, 2,4-DIHYDROXYBENZOATE AND 2-HYDROXY-4-AMINOBENZOATE AND OF THE TRY222ALA MUTANT, COMPLEXED WITH 2-HYDROXY-4-AMINOBENZOATE. EVIDENCE FOR A PROTON CHANNEL AND A NEW BINDING MODE OF THE FLAVIN RING== |
| The crystal structures of wild-type p-hydroxybenzoate hydroxylase from, Pseudomonas fluorescens, complexed with the substrate analogues, 4-aminobenzoate, 2,4-dihydroxybenzoate, and 2-hydroxy-4-aminobenzoate have, been determined at 2.3-, 2.5-, and 2.8-A resolution, respectively. In, addition, the crystal structure of a Tyr222Ala mutant, complexed with, 2-hydroxy-4-aminobenzoate, has been determined at 2.7-A resolution. The, structures have been refined to R factors between 14.5% and 15.8% for data, between 8.0 A and the high-resolution limit. The differences between these, complexes and the wild-type enzyme-substrate complex are all concentrated, in the active site region. Binding of substrate analogues bearing a, 4-amino group (4-aminobenzoate and 2-hydroxy-4-aminobenzoate) leads to, binding of a water molecule next to the active site Tyr385. As a result, a, continuous hydrogen-bonding network is present between the 4-amino group, of the substrate analogue and the side chain of His72. It is likely that, this hydrogen-bonding network is transiently present during normal, catalysis, where it may or may not function as a proton channel assisting, the deprotonation of the 4-hydroxyl group of the normal substrate upon, binding to the active site. Binding of substrate analogues bearing a, hydroxyl group at the 2-position (2,4-dihydroxybenzoate and, 2-hydroxy-4-aminobenzoate) leads to displacement of the flavin ring from, the active site. The flavin is no longer in the active site (the "in", conformation) but is in the cleft leading to the active site instead (the, "out" conformation). It is proposed that movement of the FAD out of the, active site may provide an entrance for the substrate to enter the active, site and an exit for the product to leave.
| | <StructureSection load='1pbd' size='340' side='right'caption='[[1pbd]], [[Resolution|resolution]] 2.30Å' scene=''> |
| | == Structural highlights == |
| | <table><tr><td colspan='2'>[[1pbd]] is a 1 chain structure with sequence from [https://en.wikipedia.org/wiki/Pseudomonas_fluorescens Pseudomonas fluorescens]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1PBD OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=1PBD FirstGlance]. <br> |
| | </td></tr><tr id='method'><td class="sblockLbl"><b>[[Empirical_models|Method:]]</b></td><td class="sblockDat" id="methodDat">X-ray diffraction, [[Resolution|Resolution]] 2.3Å</td></tr> |
| | <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=PAB:4-AMINOBENZOIC+ACID'>PAB</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=1pbd FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=1pbd OCA], [https://pdbe.org/1pbd PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=1pbd RCSB], [https://www.ebi.ac.uk/pdbsum/1pbd PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=1pbd ProSAT]</span></td></tr> |
| | </table> |
| | == Function == |
| | [https://www.uniprot.org/uniprot/PHHY_PSEFL PHHY_PSEFL] |
| | == 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/pb/1pbd_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=1pbd ConSurf]. |
| | <div style="clear:both"></div> |
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| ==About this Structure== | | ==See Also== |
| 1PBD is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Pseudomonas_fluorescens Pseudomonas fluorescens] with FAD and PAB as [http://en.wikipedia.org/wiki/ligands ligands]. Active as [http://en.wikipedia.org/wiki/4-hydroxybenzoate_3-monooxygenase 4-hydroxybenzoate 3-monooxygenase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=1.14.13.2 1.14.13.2] Full crystallographic information is available from [http://ispc.weizmann.ac.il/oca-bin/ocashort?id=1PBD OCA].
| | *[[Hydroxylases 3D structures|Hydroxylases 3D structures]] |
| | | __TOC__ |
| ==Reference==
| | </StructureSection> |
| Crystal structures of wild-type p-hydroxybenzoate hydroxylase complexed with 4-aminobenzoate,2,4-dihydroxybenzoate, and 2-hydroxy-4-aminobenzoate and of the Tyr222Ala mutant complexed with 2-hydroxy-4-aminobenzoate. Evidence for a proton channel and a new binding mode of the flavin ring., Schreuder HA, Mattevi A, Obmolova G, Kalk KH, Hol WG, van der Bolt FJ, van Berkel WJ, Biochemistry. 1994 Aug 23;33(33):10161-70. PMID:[http://ispc.weizmann.ac.il//pmbin/getpm?pmid=7520279 7520279]
| | [[Category: Large Structures]] |
| [[Category: 4-hydroxybenzoate 3-monooxygenase]] | |
| [[Category: Pseudomonas fluorescens]] | | [[Category: Pseudomonas fluorescens]] |
| [[Category: Single protein]]
| | [[Category: Hol WGJ]] |
| [[Category: Hol, W.G.J.]] | | [[Category: Mattevi A]] |
| [[Category: Mattevi, A.]] | | [[Category: Schreuder HA]] |
| [[Category: Schreuder, H.A.]] | |
| [[Category: FAD]]
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| [[Category: PAB]]
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| [[Category: oxidoreductase]]
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| ''Page seeded by [http://ispc.weizmann.ac.il/oca OCA ] on Tue Nov 20 23:39:42 2007''
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