5o8q

From Proteopedia

(Difference between revisions)
Jump to: navigation, search
m (Protected "5o8q" [edit=sysop:move=sysop])
Line 1: Line 1:
-
'''Unreleased structure'''
 
-
The entry 5o8q is ON HOLD until Paper Publication
+
==Crystal structure of R. ruber ADH-A, mutant Y294F, W295A==
 +
<StructureSection load='5o8q' size='340' side='right' caption='[[5o8q]], [[Resolution|resolution]] 2.22&Aring;' scene=''>
 +
== Structural highlights ==
 +
<table><tr><td colspan='2'>[[5o8q]] is a 8 chain structure. 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>
 +
</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>
 +
<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>
 +
</table>
 +
<div style="background-color:#fffaf0;">
 +
== 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.
-
Authors: Dobritzsch, D., Maurer, D., Hamnevik, E., Enugala, T.R., Widersten, M.
+
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<ref>PMID:28963762</ref>
-
Description: Crystal structure of R. ruber ADH-A, mutant Y294F, W295A
+
From MEDLINE&reg;/PubMed&reg;, a database of the U.S. National Library of Medicine.<br>
-
[[Category: Unreleased Structures]]
+
</div>
 +
<div class="pdbe-citations 5o8q" style="background-color:#fffaf0;"></div>
 +
== References ==
 +
<references/>
 +
__TOC__
 +
</StructureSection>
 +
[[Category: Dobritzsch, D]]
 +
[[Category: Enugala, T R]]
[[Category: Hamnevik, E]]
[[Category: Hamnevik, E]]
-
[[Category: Widersten, M]]
 
-
[[Category: Enugala, T.R]]
 
[[Category: Maurer, D]]
[[Category: Maurer, D]]
-
[[Category: Dobritzsch, D]]
+
[[Category: Widersten, M]]
 +
[[Category: Alcohol dehydrogenase mutant variant]]
 +
[[Category: Nadh-dependent]]
 +
[[Category: Oxidoreductase]]
 +
[[Category: Rossmann fold]]
 +
[[Category: Zn2+-dependent]]

Revision as of 07:00, 11 October 2017

Crystal structure of R. ruber ADH-A, mutant Y294F, W295A

5o8q, resolution 2.22Å

Drag the structure with the mouse to rotate

Proteopedia Page Contributors and Editors (what is this?)

OCA

Personal tools