View source for 4i5f


==Crystal structure of Ralstonia sp. alcohol dehydrogenase mutant N15G, G37D, R38V, R39S==
<StructureSection load='4i5f' size='340' side='right'caption='[[4i5f]], [[Resolution|resolution]] 2.10&Aring;' scene=''>
== Structural highlights ==
<table><tr><td colspan='2'>[[4i5f]] 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=4I5F OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=4I5F 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]] 2.1&#8491;</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=4i5f FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=4i5f OCA], [https://pdbe.org/4i5f PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=4i5f RCSB], [https://www.ebi.ac.uk/pdbsum/4i5f PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=4i5f ProSAT]</span></td></tr>
</table>
== Function ==
[https://www.uniprot.org/uniprot/C0IR58_9RALS C0IR58_9RALS] 
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== 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&reg;/PubMed&reg;, a database of the U.S. National Library of Medicine.<br>
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== References ==
<references/>
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</StructureSection>
[[Category: Large Structures]]
[[Category: Ralstonia sp. DSMZ 6428]]
[[Category: Jarasch A]]
[[Category: Lerchner A]]
[[Category: Meining W]]
[[Category: Schiefner A]]
[[Category: Skerra A]]