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| [[Image:1kbo.gif|left|200px]]<br />
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| <applet load="1kbo" size="450" color="white" frame="true" align="right" spinBox="true"
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| caption="1kbo, resolution 2.30Å" />
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| '''Complex of Human recombinant NAD(P)H:Quinone Oxide reductase type 1 with 5-methoxy-1,2-dimethyl-3-(phenoxymethyl)indole-4,7-dione (ES1340)'''<br />
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| ==Overview== | | ==Complex of Human recombinant NAD(P)H:Quinone Oxide reductase type 1 with 5-methoxy-1,2-dimethyl-3-(phenoxymethyl)indole-4,7-dione (ES1340)== |
| We report the characterization of, 5-methoxy-1,2-dimethyl-3-[(4-nitrophenoxy)methyl]indole-4,7-dione (ES936), as a mechanism-based inhibitor of NQO1. Inactivation of NQO1 by ES936 was, time- and concentration-dependent and required the presence of a pyridine, nucleotide cofactor consistent with a need for metabolic activation. That, ES936 was an efficient inhibitor was demonstrated in these studies by the, low partition ratio (1.40 +/- 0.03). The orientation of ES936 in the, active site of NQO1 was examined by X-ray crystallography and found to be, opposite to that observed for other indolequinones acting as substrates., ES936 was oriented in such a manner that, after enzymatic reduction and, loss of a nitrophenol leaving group, a reactive iminium species was, located in close proximity to nucleophilic His 162 and Tyr 127 and Tyr 129, residues in the active site. To determine if ES936 was covalently, modifying NQO1, ES936-treated protein was analyzed by electrospray, ionization liquid chromatography/mass spectrometry (ESI-LC/MS). The, control NQO1 protein had a mass of 30864 +/- 6 Da (n = 20, theoretical, 30868.6 Da) which increased by 217 Da after ES936 treatment (31081 +/- 7, Da, n = 20) in the presence of NADH. The shift in mass was consistent with, adduction of NQO1 by the reactive iminium derived from ES936 (M + 218 Da)., Chymotryptic digestion of the protein followed by LC/MS analysis located a, tetrapeptide spanning amino acids 126-129 which was adducted with the, reactive iminium species derived from ES936. LC/MS/MS analysis of the, peptide fragment confirmed adduction of either Tyr 127 or Tyr 129, residues. This work demonstrates that ES936 is a potent mechanism-based, inhibitor of NQO1 and may be a useful tool in defining the role of NQO1 in, cellular systems and in vivo.
| | <StructureSection load='1kbo' size='340' side='right'caption='[[1kbo]], [[Resolution|resolution]] 2.30Å' scene=''> |
| | == Structural highlights == |
| | <table><tr><td colspan='2'>[[1kbo]] is a 4 chain structure with sequence from [https://en.wikipedia.org/wiki/Homo_sapiens Homo sapiens]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1KBO OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=1KBO 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=340:5-METHOXY-1,2-DIMETHYL-3-(PHENOXYMETHYL)INDOLE-4,7-DIONE'>340</scene>, <scene name='pdbligand=FAD:FLAVIN-ADENINE+DINUCLEOTIDE'>FAD</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=1kbo FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=1kbo OCA], [https://pdbe.org/1kbo PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=1kbo RCSB], [https://www.ebi.ac.uk/pdbsum/1kbo PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=1kbo ProSAT]</span></td></tr> |
| | </table> |
| | == Function == |
| | [https://www.uniprot.org/uniprot/NQO1_HUMAN NQO1_HUMAN] The enzyme apparently serves as a quinone reductase in connection with conjugation reactions of hydroquinons involved in detoxification pathways as well as in biosynthetic processes such as the vitamin K-dependent gamma-carboxylation of glutamate residues in prothrombin synthesis. |
| | == 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/kb/1kbo_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=1kbo ConSurf]. |
| | <div style="clear:both"></div> |
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| ==Disease== | | ==See Also== |
| Known diseases associated with this structure: Benzene toxicity, susceptibility to OMIM:[[http://www.ncbi.nlm.nih.gov/entrez/dispomim.cgi?id=125860 125860]], Leukemia, post-chemotherapy, susceptibility to OMIM:[[http://www.ncbi.nlm.nih.gov/entrez/dispomim.cgi?id=125860 125860]]
| | *[[Quinone reductase 3D structures|Quinone reductase 3D structures]] |
| | | __TOC__ |
| ==About this Structure==
| | </StructureSection> |
| 1KBO is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Homo_sapiens Homo sapiens] with 340 and FAD as [http://en.wikipedia.org/wiki/ligands ligands]. Active as [http://en.wikipedia.org/wiki/NAD(P)H_dehydrogenase_(quinone) NAD(P)H dehydrogenase (quinone)], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=1.6.5.2 1.6.5.2] Full crystallographic information is available from [http://ispc.weizmann.ac.il/oca-bin/ocashort?id=1KBO OCA].
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| ==Reference==
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| Characterization of a mechanism-based inhibitor of NAD(P)H:quinone oxidoreductase 1 by biochemical, X-ray crystallographic, and mass spectrometric approaches., Winski SL, Faig M, Bianchet MA, Siegel D, Swann E, Fung K, Duncan MW, Moody CJ, Amzel LM, Ross D, Biochemistry. 2001 Dec 18;40(50):15135-42. PMID:[http://ispc.weizmann.ac.il//pmbin/getpm?pmid=11735396 11735396]
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| [[Category: Homo sapiens]] | | [[Category: Homo sapiens]] |
| [[Category: NAD(P)H dehydrogenase (quinone)]] | | [[Category: Large Structures]] |
| [[Category: Single protein]]
| | [[Category: Amzel LM]] |
| [[Category: Amzel, L.M.]] | | [[Category: Bianchet MA]] |
| [[Category: Bianchet, M.A.]] | | [[Category: Faig M]] |
| [[Category: Faig, M.]] | |
| [[Category: 340]]
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| [[Category: FAD]]
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| [[Category: flavoenzyme]]
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| [[Category: prodrug-enzyme complex]]
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| ''Page seeded by [http://ispc.weizmann.ac.il/oca OCA ] on Mon Nov 12 17:49:47 2007''
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