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4i5f

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Crystal structure of Ralstonia sp. alcohol dehydrogenase mutant N15G, G37D, R38V, R39S

Structural highlights

4i5f is a 8 chain structure with sequence from Ralstonia sp. DSMZ 6428. Full crystallographic information is available from OCA. For a guided tour on the structure components use FirstGlance.
Method:	X-ray diffraction, Resolution 2.1Å
Resources:	FirstGlance, OCA, PDBe, RCSB, PDBsum, ProSAT

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

1 Structural highlights
2 Function
3 Publication Abstract from PubMed
4 References

Proteopedia Page Contributors and Editors (what is this?)

OCA

Retrieved from "http://52.214.119.220/wiki/index.php/4i5f"

@@ Line 1: / Line 1: @@
 ==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=''>
+<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 [http://en.wikipedia.org/wiki/Saccharomyces_mikatae Saccharomyces mikatae]. 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 [http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=4I5F FirstGlance]. <br>
+<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='related'><td class="sblockLbl"><b>[[Related_structure|Related:]]</b></td><td class="sblockDat">[[4i5d|4i5d]], [[4i5e|4i5e]], [[4i5g|4i5g]]</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]] 2.1&#8491;</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=114525 Saccharomyces mikatae])</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>
-<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=4i5f FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=4i5f OCA], [http://www.rcsb.org/pdb/explore.do?structureId=4i5f RCSB], [http://www.ebi.ac.uk/pdbsum/4i5f PDBsum]</span></td></tr>
 </table>
+== Function ==
+[https://www.uniprot.org/uniprot/C0IR58_9RALS C0IR58_9RALS]
 <div style="background-color:#fffaf0;">
 == Publication Abstract from PubMed ==
@@ Line 15: / Line 17: @@
 From MEDLINE&reg;/PubMed&reg;, a database of the U.S. National Library of Medicine.<br>
 </div>
+<div class="pdbe-citations 4i5f" style="background-color:#fffaf0;"></div>
 == References ==
 <references/>
 __TOC__
 </StructureSection>
-[[Category: Saccharomyces mikatae]]
+[[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]]

4i5f

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Crystal structure of Ralstonia sp. alcohol dehydrogenase mutant N15G, G37D, R38V, R39S

Structural highlights

Function

Publication Abstract from PubMed

References

Contents

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