4i5g
From Proteopedia
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| - | + | ==Crystal structure of Ralstonia sp. alcohol dehydrogenase mutant N15G, G37D, R38V, R39S, A86N, S88A== | |
| - | + | <StructureSection load='4i5g' size='340' side='right' caption='[[4i5g]], [[Resolution|resolution]] 2.30Å' scene=''> | |
| - | + | == Structural highlights == | |
| + | <table><tr><td colspan='2'>[[4i5g]] 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=4I5G OCA]. For a <b>guided tour on the structure components</b> use [http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=4I5G FirstGlance]. <br> | ||
| + | </td></tr><tr id='related'><td class="sblockLbl"><b>[[Related_structure|Related:]]</b></td><td class="sblockDat">[[4i5e|4i5e]], [[4i5f|4i5f]]</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'>[http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=4i5g FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=4i5g OCA], [http://www.rcsb.org/pdb/explore.do?structureId=4i5g RCSB], [http://www.ebi.ac.uk/pdbsum/4i5g PDBsum]</span></td></tr> | ||
| + | </table> | ||
| + | <div style="background-color:#fffaf0;"> | ||
| + | == 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<ref>PMID:23686719</ref> | |
| - | + | ||
| - | == | + | From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br> |
| - | + | </div> | |
| + | == References == | ||
| + | <references/> | ||
| + | __TOC__ | ||
| + | </StructureSection> | ||
[[Category: Ralstonia sp. dsmz 6428]] | [[Category: Ralstonia sp. dsmz 6428]] | ||
| - | [[Category: Jarasch, A | + | [[Category: Jarasch, A]] |
| - | [[Category: Lerchner, A | + | [[Category: Lerchner, A]] |
| - | [[Category: Meining, W | + | [[Category: Meining, W]] |
| - | [[Category: Schiefner, A | + | [[Category: Schiefner, A]] |
| - | [[Category: Skerra, A | + | [[Category: Skerra, A]] |
[[Category: Alcohol dehydrogenase]] | [[Category: Alcohol dehydrogenase]] | ||
[[Category: Cosubstrate specificity]] | [[Category: Cosubstrate specificity]] | ||
[[Category: Nadh]] | [[Category: Nadh]] | ||
[[Category: Oxidoreductase]] | [[Category: Oxidoreductase]] | ||
| - | [[Category: Ralstonia sp | + | [[Category: Ralstonia sp]] |
[[Category: Rasadh]] | [[Category: Rasadh]] | ||
[[Category: Rossmann fold]] | [[Category: Rossmann fold]] | ||
[[Category: S-phenylethanol]] | [[Category: S-phenylethanol]] | ||
[[Category: Short-chain-dehydrogenases/reductase]] | [[Category: Short-chain-dehydrogenases/reductase]] | ||
Revision as of 12:31, 21 December 2014
Crystal structure of Ralstonia sp. alcohol dehydrogenase mutant N15G, G37D, R38V, R39S, A86N, S88A
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