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| | <StructureSection load='5iwe' size='340' side='right'caption='[[5iwe]], [[Resolution|resolution]] 1.71Å' scene=''> | | <StructureSection load='5iwe' size='340' side='right'caption='[[5iwe]], [[Resolution|resolution]] 1.71Å' scene=''> |
| | == Structural highlights == | | == Structural highlights == |
| - | <table><tr><td colspan='2'>[[5iwe]] is a 1 chain structure. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=5IWE OCA]. For a <b>guided tour on the structure components</b> use [http://proteopedia.org/fgij/fg.htm?mol=5IWE FirstGlance]. <br> | + | <table><tr><td colspan='2'>[[5iwe]] 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=5IWE OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=5IWE FirstGlance]. <br> |
| - | </td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=EDO:1,2-ETHANEDIOL'>EDO</scene>, <scene name='pdbligand=GOL:GLYCEROL'>GOL</scene>, <scene name='pdbligand=PEG:DI(HYDROXYETHYL)ETHER'>PEG</scene>, <scene name='pdbligand=W81:(5R,6R)-6-AZANIUMYL-5-ETHOXYCYCLOHEXA-1,3-DIENE-1-CARBOXYLATE'>W81</scene></td></tr> | + | </td></tr><tr id='method'><td class="sblockLbl"><b>[[Empirical_models|Method:]]</b></td><td class="sblockDat" id="methodDat">X-ray diffraction, [[Resolution|Resolution]] 1.71Å</td></tr> |
| - | <tr id='activity'><td class="sblockLbl"><b>Activity:</b></td><td class="sblockDat"><span class='plainlinks'>[http://en.wikipedia.org/wiki/Trans-2,3-dihydro-3-hydroxyanthranilate_isomerase Trans-2,3-dihydro-3-hydroxyanthranilate isomerase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=5.3.3.17 5.3.3.17] </span></td></tr>
| + | <tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=EDO:1,2-ETHANEDIOL'>EDO</scene>, <scene name='pdbligand=GOL:GLYCEROL'>GOL</scene>, <scene name='pdbligand=PEG:DI(HYDROXYETHYL)ETHER'>PEG</scene>, <scene name='pdbligand=W81:(5R,6R)-6-AZANIUMYL-5-ETHOXYCYCLOHEXA-1,3-DIENE-1-CARBOXYLATE'>W81</scene></td></tr> |
| - | <tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[http://proteopedia.org/fgij/fg.htm?mol=5iwe FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=5iwe OCA], [http://pdbe.org/5iwe PDBe], [http://www.rcsb.org/pdb/explore.do?structureId=5iwe RCSB], [http://www.ebi.ac.uk/pdbsum/5iwe PDBsum], [http://prosat.h-its.org/prosat/prosatexe?pdbcode=5iwe ProSAT]</span></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=5iwe FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=5iwe OCA], [https://pdbe.org/5iwe PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=5iwe RCSB], [https://www.ebi.ac.uk/pdbsum/5iwe PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=5iwe ProSAT]</span></td></tr> |
| | </table> | | </table> |
| | == Function == | | == Function == |
| - | [[http://www.uniprot.org/uniprot/PHZF_PSEFL PHZF_PSEFL]] Isomerase that catalyzes the condensation of two molecules of trans-2,3-dihydro-3-hydroxyanthranilic acid (DHHA) into the phenazine ring system. The final product is not yet known.<ref>PMID:19053436</ref> <ref>PMID:15545603</ref> | + | [https://www.uniprot.org/uniprot/PHZF_PSEFL PHZF_PSEFL] Isomerase that catalyzes the condensation of two molecules of trans-2,3-dihydro-3-hydroxyanthranilic acid (DHHA) into the phenazine ring system. The final product is not yet known.<ref>PMID:19053436</ref> <ref>PMID:15545603</ref> |
| | <div style="background-color:#fffaf0;"> | | <div style="background-color:#fffaf0;"> |
| | == Publication Abstract from PubMed == | | == Publication Abstract from PubMed == |
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| | </StructureSection> | | </StructureSection> |
| | [[Category: Large Structures]] | | [[Category: Large Structures]] |
| - | [[Category: Trans-2,3-dihydro-3-hydroxyanthranilate isomerase]] | + | [[Category: Pseudomonas fluorescens]] |
| - | [[Category: Blankenfeldt, W]] | + | [[Category: Blankenfeldt W]] |
| - | [[Category: Diederich, C]] | + | [[Category: Diederich C]] |
| - | [[Category: Complex]]
| + | |
| - | [[Category: Isomerase]]
| + | |
| - | [[Category: Substrate analogue]]
| + | |
| Structural highlights
Function
PHZF_PSEFL Isomerase that catalyzes the condensation of two molecules of trans-2,3-dihydro-3-hydroxyanthranilic acid (DHHA) into the phenazine ring system. The final product is not yet known.[1] [2]
Publication Abstract from PubMed
Phenazines are bacterial virulence and survival factors with important roles in infectious disease. PhzF catalyzes a key reaction in their biosynthesis by isomerizing (2 S,3 S)-2,3-dihydro-3-hydroxy anthranilate (DHHA) in two steps, a [1,5]-hydrogen shift followed by tautomerization to an aminoketone. While the [1,5]-hydrogen shift requires the conserved glutamate E45, suggesting acid/base catalysis, it also shows hallmarks of a sigmatropic rearrangement, namely the suprafacial migration of a non-acidic proton. To discriminate these mechanistic alternatives, we employed enzyme kinetic measurements and computational methods. Quantum mechanics/molecular mechanics (QM/MM) calculations revealed that the activation barrier of a proton shuttle mechanism involving E45 is significantly lower than that of a sigmatropic [1,5]-hydrogen shift. QM/MM also predicted a large kinetic isotope effect, which was indeed observed with deuterated substrate. For the tautomerization, QM/MM calculations suggested involvement of E45 and an active site water molecule, explaining the observed stereochemistry. Because these findings imply that PhzF can act only on a limited substrate spectrum, we also investigated the turnover of DHHA derivatives, of which only O-methyl and O-ethyl DHHA were converted. Together, these data reveal how PhzF orchestrates a water-free with a water-dependent step. Its unique mechanism, specificity and essential role in phenazine biosynthesis may offer opportunities for inhibitor development.
Mechanisms and Specificity of Phenazine Biosynthesis Protein PhzF.,Diederich C, Leypold M, Culka M, Weber H, Breinbauer R, Ullmann GM, Blankenfeldt W Sci Rep. 2017 Jul 24;7(1):6272. doi: 10.1038/s41598-017-06278-w. PMID:28740244[3]
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.
References
- ↑ Ahuja EG, Janning P, Mentel M, Graebsch A, Breinbauer R, Hiller W, Costisella B, Thomashow LS, Mavrodi DV, Blankenfeldt W. PhzA/B catalyzes the formation of the tricycle in phenazine biosynthesis. J Am Chem Soc. 2008 Dec 17;130(50):17053-61. PMID:19053436 doi:10.1021/ja806325k
- ↑ Blankenfeldt W, Kuzin AP, Skarina T, Korniyenko Y, Tong L, Bayer P, Janning P, Thomashow LS, Mavrodi DV. Structure and function of the phenazine biosynthetic protein PhzF from Pseudomonas fluorescens. Proc Natl Acad Sci U S A. 2004 Nov 23;101(47):16431-6. Epub 2004 Nov 15. PMID:15545603
- ↑ Diederich C, Leypold M, Culka M, Weber H, Breinbauer R, Ullmann GM, Blankenfeldt W. Mechanisms and Specificity of Phenazine Biosynthesis Protein PhzF. Sci Rep. 2017 Jul 24;7(1):6272. doi: 10.1038/s41598-017-06278-w. PMID:28740244 doi:http://dx.doi.org/10.1038/s41598-017-06278-w
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