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| ==Crystal structure of R. ruber ADH-A, mutant Y294F, W295A== | | ==Crystal structure of R. ruber ADH-A, mutant Y294F, W295A== |
- | <StructureSection load='5o8q' size='340' side='right' caption='[[5o8q]], [[Resolution|resolution]] 2.22Å' scene=''> | + | <StructureSection load='5o8q' size='340' side='right'caption='[[5o8q]], [[Resolution|resolution]] 2.22Å' scene=''> |
| == Structural highlights == | | == Structural highlights == |
- | <table><tr><td colspan='2'>[[5o8q]] is a 8 chain structure with sequence from [http://en.wikipedia.org/wiki/Rhodococcus_sp._m8 Rhodococcus sp. m8]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=5O8Q OCA]. For a <b>guided tour on the structure components</b> use [http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=5O8Q FirstGlance]. <br> | + | <table><tr><td colspan='2'>[[5o8q]] is a 8 chain structure with sequence from [https://en.wikipedia.org/wiki/Rhodococcus_sp._M8 Rhodococcus sp. M8]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=5O8Q OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=5O8Q FirstGlance]. <br> |
- | </td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat"><scene name='pdbligand=NAD:NICOTINAMIDE-ADENINE-DINUCLEOTIDE'>NAD</scene>, <scene name='pdbligand=ZN:ZINC+ION'>ZN</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]] 2.22Å</td></tr> |
- | <tr id='gene'><td class="sblockLbl"><b>[[Gene|Gene:]]</b></td><td class="sblockDat">BKE56_025765 ([http://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?mode=Info&srchmode=5&id=1925550 Rhodococcus sp. M8])</td></tr> | + | <tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=NAD:NICOTINAMIDE-ADENINE-DINUCLEOTIDE'>NAD</scene>, <scene name='pdbligand=ZN:ZINC+ION'>ZN</scene></td></tr> |
- | <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=5o8q FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=5o8q OCA], [http://pdbe.org/5o8q PDBe], [http://www.rcsb.org/pdb/explore.do?structureId=5o8q RCSB], [http://www.ebi.ac.uk/pdbsum/5o8q PDBsum], [http://prosat.h-its.org/prosat/prosatexe?pdbcode=5o8q 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=5o8q FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=5o8q OCA], [https://pdbe.org/5o8q PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=5o8q RCSB], [https://www.ebi.ac.uk/pdbsum/5o8q PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=5o8q ProSAT]</span></td></tr> |
| </table> | | </table> |
| + | == Function == |
| + | [https://www.uniprot.org/uniprot/A0A1Q8I6M1_9NOCA A0A1Q8I6M1_9NOCA] |
| <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 5o8q" style="background-color:#fffaf0;"></div> | | <div class="pdbe-citations 5o8q" style="background-color:#fffaf0;"></div> |
| + | |
| + | ==See Also== |
| + | *[[Alcohol dehydrogenase 3D structures|Alcohol dehydrogenase 3D structures]] |
| == References == | | == References == |
| <references/> | | <references/> |
| __TOC__ | | __TOC__ |
| </StructureSection> | | </StructureSection> |
- | [[Category: Rhodococcus sp. m8]] | + | [[Category: Large Structures]] |
- | [[Category: Dobritzsch, D]] | + | [[Category: Rhodococcus sp. M8]] |
- | [[Category: Enugala, T R]] | + | [[Category: Dobritzsch D]] |
- | [[Category: Hamnevik, E]] | + | [[Category: Enugala TR]] |
- | [[Category: Maurer, D]] | + | [[Category: Hamnevik E]] |
- | [[Category: Widersten, M]] | + | [[Category: Maurer D]] |
- | [[Category: Alcohol dehydrogenase mutant variant]]
| + | [[Category: Widersten M]] |
- | [[Category: Nadh-dependent]]
| + | |
- | [[Category: Oxidoreductase]]
| + | |
- | [[Category: Rossmann fold]]
| + | |
- | [[Category: Zn2+-dependent]]
| + | |
| Structural highlights
Function
A0A1Q8I6M1_9NOCA
Publication Abstract from PubMed
Alcohol dehydrogenase A (ADH-A) from Rhodococcus ruber DSM 44541 is a promising biocatalyst for redox transformations of arylsubstituted sec-alcohols and ketones. The enzyme is stereoselective in the oxidation of 1-phenylethanol with a 300-fold preference for the (S)-enantiomer. The low catalytic efficiency with (R)-1-phenylethanol has been attributed to nonproductive binding of this substrate at the active site. Aiming to modify the enantioselectivity, to rather favor the (R)-alcohol, and also test the possible involvement of nonproductive substrate binding as a mechanism in substrate discrimination, we performed directed laboratory evolution of ADH-A. Three targeted sites that contribute to the active-site cavity were exposed to saturation mutagenesis in a stepwise manner and the generated variants were selected for improved catalytic activity with (R)-1-phenylethanol. After three subsequent rounds of mutagenesis, selection and structure-function analysis of isolated ADH-A variants, we conclude: (1) W295 has a key role as a structural determinant in the discrimination between (R)- and (S)-1-phenylethanol and a W295A substitution fundamentally changes the stereoselectivity of the protein. One observable effect is a faster rate of NADH release, which changes the rate-limiting step of the catalytic cycle from coenzyme release to hydride transfer. (2) The obtained change in enantiopreference, from the (S)- to the (R)-alcohol, can be partly explained by a shift in the nonproductive substrate binding modes. This article is protected by copyright. All rights reserved.
Relaxation of nonproductive binding and increased rate of coenzyme release in an alcohol dehydrogenase increases turnover with a non-preferred alcohol enantiomer.,Hamnevik E, Enugala TR, Maurer D, Ntuku S, Oliveira A, Dobritzsch D, Widersten M FEBS J. 2017 Sep 30. doi: 10.1111/febs.14279. PMID:28963762[1]
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.
See Also
References
- ↑ Hamnevik E, Enugala TR, Maurer D, Ntuku S, Oliveira A, Dobritzsch D, Widersten M. Relaxation of nonproductive binding and increased rate of coenzyme release in an alcohol dehydrogenase increases turnover with a non-preferred alcohol enantiomer. FEBS J. 2017 Sep 30. doi: 10.1111/febs.14279. PMID:28963762 doi:http://dx.doi.org/10.1111/febs.14279
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