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| | <StructureSection load='7e8q' size='340' side='right'caption='[[7e8q]], [[Resolution|resolution]] 2.30Å' scene=''> | | <StructureSection load='7e8q' size='340' side='right'caption='[[7e8q]], [[Resolution|resolution]] 2.30Å' scene=''> |
| | == Structural highlights == | | == Structural highlights == |
| - | <table><tr><td colspan='2'>[[7e8q]] is a 4 chain structure with sequence from [https://en.wikipedia.org/wiki/Atcc_27511 Atcc 27511]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=7E8Q OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=7E8Q FirstGlance]. <br> | + | <table><tr><td colspan='2'>[[7e8q]] is a 4 chain structure with sequence from [https://en.wikipedia.org/wiki/Ralstonia_pickettii Ralstonia pickettii]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=7E8Q OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=7E8Q FirstGlance]. <br> |
| - | </td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=FDA:DIHYDROFLAVINE-ADENINE+DINUCLEOTIDE'>FDA</scene>, <scene name='pdbligand=NPO:P-NITROPHENOL'>NPO</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]] 2.3Å</td></tr> |
| - | <tr id='related'><td class="sblockLbl"><b>[[Related_structure|Related:]]</b></td><td class="sblockDat"><div style='overflow: auto; max-height: 3em;'>[[6jhm|6jhm]], [[7e8p|7e8p]]</div></td></tr> | + | <tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=FDA:DIHYDROFLAVINE-ADENINE+DINUCLEOTIDE'>FDA</scene>, <scene name='pdbligand=NPO:P-NITROPHENOL'>NPO</scene></td></tr> |
| - | <tr id='gene'><td class="sblockLbl"><b>[[Gene|Gene:]]</b></td><td class="sblockDat">hadA ([https://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?mode=Info&srchmode=5&id=329 ATCC 27511])</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=7e8q FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=7e8q OCA], [https://pdbe.org/7e8q PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=7e8q RCSB], [https://www.ebi.ac.uk/pdbsum/7e8q PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=7e8q 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=7e8q FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=7e8q OCA], [https://pdbe.org/7e8q PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=7e8q RCSB], [https://www.ebi.ac.uk/pdbsum/7e8q PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=7e8q ProSAT]</span></td></tr> |
| | </table> | | </table> |
| | + | == Function == |
| | + | [https://www.uniprot.org/uniprot/Q53008_RALPI Q53008_RALPI] |
| | <div style="background-color:#fffaf0;"> | | <div style="background-color:#fffaf0;"> |
| | == Publication Abstract from PubMed == | | == Publication Abstract from PubMed == |
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| | </div> | | </div> |
| | <div class="pdbe-citations 7e8q" style="background-color:#fffaf0;"></div> | | <div class="pdbe-citations 7e8q" style="background-color:#fffaf0;"></div> |
| | + | |
| | + | ==See Also== |
| | + | *[[Monooxygenase 3D structures|Monooxygenase 3D structures]] |
| | == References == | | == References == |
| | <references/> | | <references/> |
| | __TOC__ | | __TOC__ |
| | </StructureSection> | | </StructureSection> |
| - | [[Category: Atcc 27511]] | |
| | [[Category: Large Structures]] | | [[Category: Large Structures]] |
| - | [[Category: Chaiyen, P]] | + | [[Category: Ralstonia pickettii]] |
| - | [[Category: Chitnumsub, P]] | + | [[Category: Chaiyen P]] |
| - | [[Category: Jaruwat, A]] | + | [[Category: Chitnumsub P]] |
| - | [[Category: Pimviriyakul, P]]
| + | [[Category: Jaruwat A]] |
| - | [[Category: Chlorophenol 4-monooxygenase]] | + | [[Category: Pimviriyakul P]] |
| - | [[Category: Flavin monooxygenase]] | + | |
| - | [[Category: Oxidoreductase]]
| + | |
| Structural highlights
Function
Q53008_RALPI
Publication Abstract from PubMed
HadA is a flavin-dependent monooxygenase catalyzing hydroxylation plus dehalogenation/denitration which is useful for biodetoxification and bio-detection. In this study, the X-ray structure of wild-type HadA (HadAWT) co-complexed with reduced FAD (FADH(-)) and 4-nitrophenol (4NP) (HadAWT-FADH(-)-4NP) was solved at 2.3 A resolution, providing the first full package (with flavin and substrate bound) structure of a monooxygenase of this type. Residues Arg101, Gln158, Arg161, Thr193, Asp254, Arg233, and Arg439 constitute a flavin binding pocket, while the 4NP binding pocket contains the aromatic sidechain of Phe206, which provides pi-pi stacking and also is a part of the hydrophobic pocket formed by Phe155, Phe286, Thr449 and Leu457. Based on site-directed mutagenesis and stopped-flow experiments, Thr193, Asp254 and His290 are important for C4a-hydroperoxyflavin formation with His290, also serving as a catalytic base for hydroxylation. We also identified a novel structural motif of quadruple pi-stacking (pi-pi-pi-pi) provided by two 4NP and two Phe441 from two subunits. This motif promotes 4NP binding in a non-productive dead-end complex which prevents C4a-hydroperoxy-FAD formation when HadA is pre-mixed with aromatic substrates. We also solved the structure of the HadAPhe441Val-FADH(-)-4NP complex at 2.3 A resolution. Although 4NP can still bind to this variant, the quadruple pi-stacking motif was disrupted. All HadAPhe441 variants lack substrate inhibition behavior, confirming that quadruple pi-stacking is a main cause of dead-end complex formation. Moreover, the activities of these HadAPhe441 variants were improved by 20%, suggesting that insights gained from the flavin-dependent monooxygenases illustrated here should be useful for future improvement of HadA's biocatalytic applications.
Structural Insights into a Flavin-dependent Dehalogenase HadA Explain Catalysis and Substrate Inhibition via Quadruple pi-stacking.,Pimviriyakul P, Jaruwat A, Chitnumsub P, Chaiyen P J Biol Chem. 2021 Jul 9:100952. doi: 10.1016/j.jbc.2021.100952. PMID:34252455[1]
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.
See Also
References
- ↑ Pimviriyakul P, Jaruwat A, Chitnumsub P, Chaiyen P. Structural Insights into a Flavin-dependent Dehalogenase HadA Explain Catalysis and Substrate Inhibition via Quadruple pi-stacking. J Biol Chem. 2021 Jul 9:100952. doi: 10.1016/j.jbc.2021.100952. PMID:34252455 doi:http://dx.doi.org/10.1016/j.jbc.2021.100952
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