7e8p: Difference between revisions

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 ==Crystal structure of a Flavin-dependent Monooxygenase HadA wild type complexed with reduced FAD and 4-nitrophenol==
-<StructureSection load='7e8p' size='340' side='right'caption='[[7e8p]]' scene=''>
+<StructureSection load='7e8p' size='340' side='right'caption='[[7e8p]], [[Resolution|resolution]] 2.30&Aring;' scene=''>
 == Structural highlights ==
-<table><tr><td colspan='2'>Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=7E8P OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=7E8P FirstGlance]. <br>
+<table><tr><td colspan='2'>[[7e8p]] is a 4 chain structure with sequence from [https://en.wikipedia.org/wiki/Ralstonia_pickettii_dtp0602 Ralstonia pickettii dtp0602]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=7E8P OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=7E8P FirstGlance]. <br>
-</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=7e8p FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=7e8p OCA], [https://pdbe.org/7e8p PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=7e8p RCSB], [https://www.ebi.ac.uk/pdbsum/7e8p PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=7e8p ProSAT]</span></td></tr>
+</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='related'><td class="sblockLbl"><b>[[Related_structure|Related:]]</b></td><td class="sblockDat"><div style='overflow: auto; max-height: 3em;'>[[6jhm|6jhm]]</div></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=1366050 Ralstonia pickettii DTP0602])</td></tr>
+<tr id='activity'><td class="sblockLbl"><b>Activity:</b></td><td class="sblockDat"><span class='plainlinks'>[https://en.wikipedia.org/wiki/4-hydroxyphenylacetate_3-monooxygenase 4-hydroxyphenylacetate 3-monooxygenase], with EC number [https://www.brenda-enzymes.info/php/result_flat.php4?ecno=1.14.14.9 1.14.14.9] </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=7e8p FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=7e8p OCA], [https://pdbe.org/7e8p PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=7e8p RCSB], [https://www.ebi.ac.uk/pdbsum/7e8p PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=7e8p ProSAT]</span></td></tr>
 </table>
+<div style="background-color:#fffaf0;">
+== 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<ref>PMID:34252455</ref>
+From MEDLINE&reg;/PubMed&reg;, a database of the U.S. National Library of Medicine.<br>
+</div>
+<div class="pdbe-citations 7e8p" style="background-color:#fffaf0;"></div>
+== References ==
+<references/>
 __TOC__
 </StructureSection>
+[[Category: 4-hydroxyphenylacetate 3-monooxygenase]]
 [[Category: Large Structures]]
-[[Category: Chaiyen P]]
+[[Category: Ralstonia pickettii dtp0602]]
-[[Category: Chitnumsub P]]
+[[Category: Chaiyen, P]]
-[[Category: Jaruwat A]]
+[[Category: Chitnumsub, P]]
-[[Category: Pimviriyakul P]]
+[[Category: Jaruwat, A]]
+[[Category: Pimviriyakul, P]]
+[[Category: Chlorophenol 4-monooxygenase]]
+[[Category: Flavin monooxygenase]]
+[[Category: Oxidoreductase]]