View source for 7x2y


==Crystal Structure of cis-4,5-dihydrodiol phthalate dehydrogenase in complex with NAD+ and 3-Hydroxybenzoate==
<StructureSection load='7x2y' size='340' side='right'caption='[[7x2y]], [[Resolution|resolution]] 2.48&Aring;' scene=''>
== Structural highlights ==
<table><tr><td colspan='2'>[[7x2y]] is a 2 chain structure with sequence from [https://en.wikipedia.org/wiki/Comamonas_testosteroni_KF-1 Comamonas testosteroni KF-1]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=7X2Y OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=7X2Y 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]] 2.48&#8491;</td></tr>
<tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=3HB:3-HYDROXYBENZOIC+ACID'>3HB</scene>, <scene name='pdbligand=GOL:GLYCEROL'>GOL</scene>, <scene name='pdbligand=NAD:NICOTINAMIDE-ADENINE-DINUCLEOTIDE'>NAD</scene>, <scene name='pdbligand=PEG:DI(HYDROXYETHYL)ETHER'>PEG</scene>, <scene name='pdbligand=PO4:PHOSPHATE+ION'>PO4</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=7x2y FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=7x2y OCA], [https://pdbe.org/7x2y PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=7x2y RCSB], [https://www.ebi.ac.uk/pdbsum/7x2y PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=7x2y ProSAT]</span></td></tr>
</table>
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== Publication Abstract from PubMed ==
Phthalate cis-4,5-dihydrodiol dehydrogenase (PhtC), the second enzyme of the phthalate catabolic pathway, catalyzes the dehydrogenation of cis-4,5-dihydrodiol phthalate (DDP). Here, we report the structural and biochemical characterization of PhtC from Comamonas testosteroni KF1 (PhtCKF1). With biochemical experiments, we have determined the enzyme's catalytic efficiency (kcat/Km) with DDP as 2.6 +/- 0.5 M(-1)s(-1), over 10-fold higher than with cis-3,4-dihydrodiol phthalate (CDP). To understand the structural basis of these reactions, the crystal structures of PhtCKF1 in apo-form, the binary complex with NAD(+), and the ternary complex with NAD(+) and 3-hydroxybenzoate (3HB) were determined. These crystal structures reveal that the binding of 3HB induces a conformational change in the substrate-binding loop. This conformational change causes the opening of the NAD (+) binding site while trapping the 3HB. The PhtCKF1 crystal structures show that the catalytic domain of PhtCKF1 is larger than that of other structurally characterized homologs and does not align with other cis-diol dehydrogenases. Structural and mutational analysis of the substrate-binding loop residues, Arg164 and Glu167 establish that conformational flexibility of this loop is necessary for positioning the substrate in a catalytically competent pose, as substitution of either of these residues to Ala did not yield the dehydrogenation activity. Further, based on the crystal structures of PhtCKF1 and related structural homologs, a reaction mechanism is proposed. Finally, with the biochemical analysis of a variant M251LPhtCKF1, the broader substrate specificity of this enzyme is explained.

Conformational flexibility enables catalysis of phthalate cis-4,5-dihydrodiol dehydrogenase.,Mahto JK, Sharma M, Neetu N, Kayastha A, Aggarwal S, Kumar P Arch Biochem Biophys. 2022 Sep 30;727:109314. doi: 10.1016/j.abb.2022.109314., Epub 2022 Jun 3. PMID:35667443<ref>PMID:35667443</ref>

From MEDLINE&reg;/PubMed&reg;, a database of the U.S. National Library of Medicine.<br>
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<div class="pdbe-citations 7x2y" style="background-color:#fffaf0;"></div>
== References ==
<references/>
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</StructureSection>
[[Category: Comamonas testosteroni KF-1]]
[[Category: Large Structures]]
[[Category: Kumar P]]
[[Category: Mahto JK]]
[[Category: Sharma M]]