View source for 5zed


==Crystal structure of Kluyveromyces polyspora ADH (KpADH) mutant (E214V/T215S)==
<StructureSection load='5zed' size='340' side='right'caption='[[5zed]], [[Resolution|resolution]] 2.20&Aring;' scene=''>
== Structural highlights ==
<table><tr><td colspan='2'>[[5zed]] is a 2 chain structure with sequence from [https://en.wikipedia.org/wiki/Vanderwaltozyma_polyspora_DSM_70294 Vanderwaltozyma polyspora DSM 70294]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=5ZED OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=5ZED 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]] 2.199&#8491;</td></tr>
<tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=NAP:NADP+NICOTINAMIDE-ADENINE-DINUCLEOTIDE+PHOSPHATE'>NAP</scene></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=5zed FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=5zed OCA], [https://pdbe.org/5zed PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=5zed RCSB], [https://www.ebi.ac.uk/pdbsum/5zed PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=5zed ProSAT]</span></td></tr>
</table>
== Function ==
[https://www.uniprot.org/uniprot/A7TM80_VANPO A7TM80_VANPO] 
<div style="background-color:#fffaf0;">
== Publication Abstract from PubMed ==
Diaryl ketones are important building blocks for synthesizing pharmaceuticals and are generally regarded as "difficult-to-reduce" ketones due to the large steric hindrance of their two bulky aromatic side chains. Alcohol dehydrogenase from Kluyveromyces polyspora ( KpADH) has been identified as a robust biocatalyst due to its high conversion of diaryl ketone substrate (4-chlorophenyl)(pyridine-2-yl)ketone (CPMK) with a moderate R-selectivity of 82% ee. To modulate the stereoselectivity of KpADH, a "polarity scanning" strategy was proposed, in which six key residues inside and at the entrance of the substrate binding pocket were identified. After iterative combinatorial mutagenesis, variants Mu-R2 and Mu-S5 with enhanced (99.2% ee, R) and inverted (97.8% ee, S) stereoselectivity were obtained. The crystal structures of KpADH and two mutants in complex with NADPH were resolved to elucidate the evolution of enantioselective inversion. Based on MD simulation, Mu-R2-CPMKProR and Mu-S5-CPMKProS were more favorable in the formation of prereaction states. Interestingly, a quadrilateral plane formed by alpha-carbons of four residues (N136, V161, C237, and G214) was identified at the entrance of the substrate binding pocket of Mu-S5; this plane acts as a "polar gate" for substrates. Due to the discrepancy in charge characteristics between chlorophenyl and pyridine substituents, the pro- S orientation of CPMK is defined when it passes through the "polar gate" in Mu-S5, whereas the similar plane in wild-type is blocked by several aromatic residues. Our result paves the way for engineering stereocomplementary ADH toward bulky diaryl ketones and provides structural insight into the mechanism of stereoselective inversion.

Structural Insight into Enantioselective Inversion of an Alcohol Dehydrogenase Reveals a "Polar Gate" in Stereorecognition of Diaryl Ketones.,Zhou J, Wang Y, Xu G, Wu L, Han R, Schwaneberg U, Rao Y, Zhao YL, Zhou J, Ni Y J Am Chem Soc. 2018 Oct 3;140(39):12645-12654. doi: 10.1021/jacs.8b08640. Epub, 2018 Sep 24. PMID:30247889<ref>PMID:30247889</ref>

From MEDLINE&reg;/PubMed&reg;, a database of the U.S. National Library of Medicine.<br>
</div>
<div class="pdbe-citations 5zed" style="background-color:#fffaf0;"></div>
== References ==
<references/>
__TOC__
</StructureSection>
[[Category: Large Structures]]
[[Category: Vanderwaltozyma polyspora DSM 70294]]
[[Category: Hou XD]]
[[Category: Ni Y]]
[[Category: Rao YJ]]
[[Category: Wang Y]]
[[Category: Wu L]]
[[Category: Xu GC]]
[[Category: ZHou JH]]
[[Category: Zhou JY]]