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| | ==Crystal structure of Ralstonia sp. alcohol dehydrogenase in its apo form== | | ==Crystal structure of Ralstonia sp. alcohol dehydrogenase in its apo form== |
| - | <StructureSection load='4i5d' size='340' side='right' caption='[[4i5d]], [[Resolution|resolution]] 2.40Å' scene=''> | + | <StructureSection load='4i5d' size='340' side='right'caption='[[4i5d]], [[Resolution|resolution]] 2.40Å' scene=''> |
| | == Structural highlights == | | == Structural highlights == |
| - | <table><tr><td colspan='2'>[[4i5d]] is a 8 chain structure with sequence from [http://en.wikipedia.org/wiki/Ralstonia_sp._dsmz_6428 Ralstonia sp. dsmz 6428]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=4I5D OCA]. For a <b>guided tour on the structure components</b> use [http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=4I5D FirstGlance]. <br> | + | <table><tr><td colspan='2'>[[4i5d]] is a 8 chain structure with sequence from [https://en.wikipedia.org/wiki/Ralstonia_sp._DSMZ_6428 Ralstonia sp. DSMZ 6428]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=4I5D OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=4I5D FirstGlance]. <br> |
| - | </td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat"><scene name='pdbligand=SO4:SULFATE+ION'>SO4</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]] 2.4Å</td></tr> |
| - | <tr id='related'><td class="sblockLbl"><b>[[Related_structure|Related:]]</b></td><td class="sblockDat">[[4i5e|4i5e]], [[4i5f|4i5f]], [[4i5g|4i5g]]</td></tr>
| + | <tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=SO4:SULFATE+ION'>SO4</scene></td></tr> |
| - | <tr id='gene'><td class="sblockLbl"><b>[[Gene|Gene:]]</b></td><td class="sblockDat">RasADH ([http://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?mode=Info&srchmode=5&id=517192 Ralstonia sp. DSMZ 6428])</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=4i5d FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=4i5d OCA], [https://pdbe.org/4i5d PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=4i5d RCSB], [https://www.ebi.ac.uk/pdbsum/4i5d PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=4i5d ProSAT]</span></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=4i5d FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=4i5d OCA], [http://pdbe.org/4i5d PDBe], [http://www.rcsb.org/pdb/explore.do?structureId=4i5d RCSB], [http://www.ebi.ac.uk/pdbsum/4i5d PDBsum], [http://prosat.h-its.org/prosat/prosatexe?pdbcode=4i5d ProSAT]</span></td></tr> | + | |
| | </table> | | </table> |
| | + | == Function == |
| | + | [https://www.uniprot.org/uniprot/C0IR58_9RALS C0IR58_9RALS] |
| | <div style="background-color:#fffaf0;"> | | <div style="background-color:#fffaf0;"> |
| | == Publication Abstract from PubMed == | | == Publication Abstract from PubMed == |
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| | __TOC__ | | __TOC__ |
| | </StructureSection> | | </StructureSection> |
| - | [[Category: Ralstonia sp. dsmz 6428]] | + | [[Category: Large Structures]] |
| - | [[Category: Jarasch, A]] | + | [[Category: Ralstonia sp. DSMZ 6428]] |
| - | [[Category: Lerchner, A]] | + | [[Category: Jarasch A]] |
| - | [[Category: Meining, W]] | + | [[Category: Lerchner A]] |
| - | [[Category: Schiefner, A]] | + | [[Category: Meining W]] |
| - | [[Category: Skerra, A]] | + | [[Category: Schiefner A]] |
| - | [[Category: Alcohol dehydrogenase]]
| + | [[Category: Skerra A]] |
| - | [[Category: Cosubstrate specificity]]
| + | |
| - | [[Category: Nadh]]
| + | |
| - | [[Category: Oxidoreductase]]
| + | |
| - | [[Category: Ralstonia sp]]
| + | |
| - | [[Category: Rasadh]]
| + | |
| - | [[Category: Rossmann fold]]
| + | |
| - | [[Category: S-phenylethanol]]
| + | |
| - | [[Category: Short-chain-dehydrogenases/reductase]]
| + | |
| Structural highlights
Function
C0IR58_9RALS
Publication Abstract from PubMed
The NADP+ -dependent alcohol dehydrogenase from Ralstonia sp. (RasADH) belongs to the protein superfamily of short-chain dehydrogenases/reductases. As an enzyme that accepts different types of substrates - including bulky-bulky as well as small-bulky secondary alcohols or ketones - with high stereoselectivity, it offers potential as a biocatalyst for industrial biotechnology. To understand substrate and cosubstrate specificities of RasADH we determined the crystal structure of the apo-enzyme as well as its NADP+ -bound state with resolutions down to 2.8 A. RasADH displays a homotetrameric quaternary structure that can be described as a dimer of homodimers while in each subunit a seven-stranded parallel beta-sheet, flanked by three alpha-helices on each side, forms a Rossmann fold-type dinucleotide binding domain. Docking of the well known substrate (S)-1-phenylethanol clearly revealed the structural determinants of stereospecificity. To favour practical RasADH application in the context of established cofactor recycling systems, for example those involving an NADH-dependent amino acid dehydrogenase, we attempted to rationally change its cosubstrate specificity from NADP+ to NAD+ utilizing the structural information that NADP+ specificity is largely conferred by the residues Asn15, Gly37, Arg38, and Arg39. Furthermore, an extensive sequence alignment with homologous dehydrogenases that have different cosubstrate specificities revealed a modified general SDR motif ASNG (instead of NNAG) at positions 86-89 of RasADH. Consequently, we constructed mutant enzymes with one (G37D), four (N15G/G37D/R38V/R39S) and six (N15G/G37D/R38V/R39S/A86N/S88A) amino acid exchanges. RasADH(N15G/G37D/R38V/R39S) was better able to accept NAD+ while showing much reduced catalytic efficiency with NADP+ , leading to a change in NADH/NADPH specificity by a factor of approximately 3.6 million. Biotechnol. Bioeng. (c) 2013 Wiley Periodicals, Inc.
Crystallographic analysis and structure-guided engineering of NADPH-dependent Ralstonia sp. alcohol dehydrogenase toward NADH cosubstrate specificity.,Lerchner A, Jarasch A, Meining W, Schiefner A, Skerra A Biotechnol Bioeng. 2013 May 18. doi: 10.1002/bit.24956. PMID:23686719[1]
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.
References
- ↑ Lerchner A, Jarasch A, Meining W, Schiefner A, Skerra A. Crystallographic analysis and structure-guided engineering of NADPH-dependent Ralstonia sp. alcohol dehydrogenase toward NADH cosubstrate specificity. Biotechnol Bioeng. 2013 May 18. doi: 10.1002/bit.24956. PMID:23686719 doi:10.1002/bit.24956
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