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| ==Structure of alcohol dehydrogenase from Drosophila lebanonesis T114V mutant complexed with NAD+== | | ==Structure of alcohol dehydrogenase from Drosophila lebanonesis T114V mutant complexed with NAD+== |
| <StructureSection load='3rj5' size='340' side='right' caption='[[3rj5]], [[Resolution|resolution]] 1.45Å' scene=''> | | <StructureSection load='3rj5' size='340' side='right'caption='[[3rj5]], [[Resolution|resolution]] 1.45Å' scene=''> |
| == Structural highlights == | | == Structural highlights == |
| <table><tr><td colspan='2'>[[3rj5]] is a 2 chain structure with sequence from [http://en.wikipedia.org/wiki/Drole Drole]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=3RJ5 OCA]. For a <b>guided tour on the structure components</b> use [http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=3RJ5 FirstGlance]. <br> | | <table><tr><td colspan='2'>[[3rj5]] is a 2 chain structure with sequence from [https://en.wikipedia.org/wiki/Scaptodrosophila_lebanonensis Scaptodrosophila lebanonensis]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=3RJ5 OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=3RJ5 FirstGlance]. <br> |
| </td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat"><scene name='pdbligand=ACY:ACETIC+ACID'>ACY</scene>, <scene name='pdbligand=NAD:NICOTINAMIDE-ADENINE-DINUCLEOTIDE'>NAD</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]] 1.45Å</td></tr> |
| <tr id='related'><td class="sblockLbl"><b>[[Related_structure|Related:]]</b></td><td class="sblockDat">[[1a4u|1a4u]], [[1b15|1b15]]</td></tr>
| | <tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=ACY:ACETIC+ACID'>ACY</scene>, <scene name='pdbligand=NAD:NICOTINAMIDE-ADENINE-DINUCLEOTIDE'>NAD</scene></td></tr> |
| <tr id='gene'><td class="sblockLbl"><b>[[Gene|Gene:]]</b></td><td class="sblockDat">Adh ([http://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?mode=Info&srchmode=5&id=7225 DROLE])</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=3rj5 FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=3rj5 OCA], [https://pdbe.org/3rj5 PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=3rj5 RCSB], [https://www.ebi.ac.uk/pdbsum/3rj5 PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=3rj5 ProSAT]</span></td></tr> |
| <tr id='activity'><td class="sblockLbl"><b>Activity:</b></td><td class="sblockDat"><span class='plainlinks'>[http://en.wikipedia.org/wiki/Alcohol_dehydrogenase Alcohol dehydrogenase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=1.1.1.1 1.1.1.1] </span></td></tr>
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| <tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=3rj5 FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=3rj5 OCA], [http://pdbe.org/3rj5 PDBe], [http://www.rcsb.org/pdb/explore.do?structureId=3rj5 RCSB], [http://www.ebi.ac.uk/pdbsum/3rj5 PDBsum], [http://prosat.h-its.org/prosat/prosatexe?pdbcode=3rj5 ProSAT]</span></td></tr> | |
| </table> | | </table> |
| <div style="background-color:#fffaf0;">
| | == Function == |
| == Publication Abstract from PubMed == | | [https://www.uniprot.org/uniprot/ADH_DROLE ADH_DROLE] |
| All drosophilid alcohol dehydrogenases contain an eight-member water chain connecting the active site with the solvent at the dimer interface. A similar water chain has also been shown to exist in other short-chain dehydrogenase/reductase (SDR) enzymes, including therapeutically important SDRs. The role of this water chain in the enzymatic reaction is unknown, but it has been proposed to be involved in a proton relay system. In the present study, a connecting link in the water chain was removed by mutating Thr114 to Val114 in Scaptodrosophila lebanonensis alcohol dehydrogenase (SlADH). This threonine is conserved in all drosophilid alcohol dehydrogenases but not in other SDRs. X-ray crystallography of the SlADH(T114V) mutant revealed a broken water chain, the overall 3D structure of the binary enzyme-NAD(+) complex was almost identical to the wild-type enzyme (SlADH(wt) ). As for the SlADH(wt) , steady-state kinetic studies revealed that catalysis by the SlADH(T114V) mutant was consistent with a compulsory ordered reaction mechanism where the co-enzyme binds to the free enzyme. The mutation caused a reduction of the k(on) velocity for NAD(+) and its binding strength to the enzyme, as well as the rate of hydride transfer (k) in the ternary enzyme-NAD(+) -alcohol complex. Furthermore, it increased the pK(a) value of the group in the binary enzyme-NAD(+) complex that regulates the k(on) velocity of alcohol and alcohol-competitive inhibitors. Overall, the results indicate that an intact water chain is essential for optimal enzyme activity and participates in a proton relay system during catalysis. Database The X-ray crystallography data are available in under Protein Data Bank accession numbers 3RJ5 (SlADH(T114V) structure determined from C2 crystals) and 3RJ9 (SlADH(T114V) structure determined from P2(1) crystals) Structured digital abstract * SlADH T114V and SlADH T114V bind by x-ray crystallography (View Interaction: 1, 2).
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| An intact eight-membered water chain in drosophilid alcohol dehydrogenases is essential for optimal enzyme activity.,Wuxiuer Y, Morgunova E, Cols N, Popov A, Karshikoff A, Sylte I, Gonzalez-Duarte R, Ladenstein R, Winberg JO FEBS J. 2012 Aug;279(16):2940-2956. doi: 10.1111/j.1742-4658.2012.08675.x. Epub, 2012 Jul 19. PMID:22741949<ref>PMID:22741949</ref>
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| From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br>
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| </div>
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| <div class="pdbe-citations 3rj5" style="background-color:#fffaf0;"></div>
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| ==See Also== | | ==See Also== |
| *[[Alcohol dehydrogenase|Alcohol dehydrogenase]] | | *[[Alcohol dehydrogenase 3D structures|Alcohol dehydrogenase 3D structures]] |
| == References ==
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| <references/>
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| __TOC__ | | __TOC__ |
| </StructureSection> | | </StructureSection> |
| [[Category: Alcohol dehydrogenase]] | | [[Category: Large Structures]] |
| [[Category: Drole]] | | [[Category: Scaptodrosophila lebanonensis]] |
| [[Category: Cols, N]] | | [[Category: Cols N]] |
| [[Category: Gonzales-Duarte, R]] | | [[Category: Gonzales-Duarte R]] |
| [[Category: Karshikoff, A]] | | [[Category: Karshikoff A]] |
| [[Category: Ladenstein, R]] | | [[Category: Ladenstein R]] |
| [[Category: Morgunova, E]] | | [[Category: Morgunova E]] |
| [[Category: Popov, A]] | | [[Category: Popov A]] |
| [[Category: Sylte, I]] | | [[Category: Sylte I]] |
| [[Category: Winberg, J O]] | | [[Category: Winberg JO]] |
| [[Category: Wuxiuer, Y]] | | [[Category: Wuxiuer Y]] |
| [[Category: Detoxification]]
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| [[Category: Nad metabolism]]
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| [[Category: Oxidoreductase]]
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| [[Category: Short-chain dehydrogenases/reductase]]
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