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==Crystal Structure of Two-Domain Laccase mutant Y230A from Streptomyces griseoflavus== | ==Crystal Structure of Two-Domain Laccase mutant Y230A from Streptomyces griseoflavus== | ||
<StructureSection load='7pen' size='340' side='right'caption='[[7pen]]' scene=''> | <StructureSection load='7pen' size='340' side='right'caption='[[7pen]], [[Resolution|resolution]] 1.60Å' scene=''> | ||
== Structural highlights == | == Structural highlights == | ||
<table><tr><td colspan='2'>Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=7PEN OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=7PEN FirstGlance]. <br> | <table><tr><td colspan='2'>[[7pen]] is a 6 chain structure with sequence from [https://en.wikipedia.org/wiki/Streptomyces_griseoflavus Streptomyces griseoflavus]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=7PEN OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=7PEN FirstGlance]. <br> | ||
</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=7pen FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=7pen OCA], [https://pdbe.org/7pen PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=7pen RCSB], [https://www.ebi.ac.uk/pdbsum/7pen PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=7pen ProSAT]</span></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]] 1.6Å</td></tr> | ||
<tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=CU:COPPER+(II)+ION'>CU</scene>, <scene name='pdbligand=NA:SODIUM+ION'>NA</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=7pen FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=7pen OCA], [https://pdbe.org/7pen PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=7pen RCSB], [https://www.ebi.ac.uk/pdbsum/7pen PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=7pen ProSAT]</span></td></tr> | |||
</table> | </table> | ||
== Function == | |||
[https://www.uniprot.org/uniprot/A0A0M4FJ81_9ACTN A0A0M4FJ81_9ACTN] | |||
<div style="background-color:#fffaf0;"> | |||
== Publication Abstract from PubMed == | |||
Laccases catalyze the oxidation of substrates with the concomitant reduction of oxygen to water. Recently, we found that polar residues located in tunnels leading to Cu2 and Cu3 ions control oxygen entrance (His 165) and proton transport (Arg 240) of two-domain laccase (2D) from Streptomyces griseoflavus (SgfSL). In this work, we have focused on optimizing the substrate-binding pocket (SBP) of SgfSL while simultaneously adjusting the oxygen reduction process. SgfSL variants with three single (Met199Ala, Met199Gly, and Tyr230Ala) and three double amino acid residues substitutions (Met199Gly/His165Ala, His165Ala/Arg240His, Met199Gly/Arg240His) were constructed, purified, and investigated. Combination of substitutions in the SBP and in the tunnel leading to Cu2 ion (Met199Gly/Arg240His) increased SgfSL catalytic activity towards ABTS by 5-fold, and towards 2.6-DMP by 16-fold. The high activity of the Met199Gly/Arg240His variant can be explained by the combined effect of the SBP geometry optimization (Met199Gly) and increased proton flux via the tunnel leading to Cu2 ion (Arg240His). Moreover, the variant with Met199Gly and His165Ala mutations did not significantly increase SgfSL's activity, but led to a drastic shift in the optimal pH of 2.6-DMP oxidation. These results indicate that His 165 not only regulates oxygen access, but it also participates in proton transport in 2D laccases. | |||
Engineering the Catalytic Properties of Two-Domain Laccase from Streptomyces griseoflavus Ac-993.,Kolyadenko I, Scherbakova A, Kovalev K, Gabdulkhakov A, Tishchenko S Int J Mol Sci. 2021 Dec 22;23(1). pii: ijms23010065. doi: 10.3390/ijms23010065. PMID:35008493<ref>PMID:35008493</ref> | |||
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br> | |||
</div> | |||
<div class="pdbe-citations 7pen" style="background-color:#fffaf0;"></div> | |||
==See Also== | |||
*[[Laccase 3D structures|Laccase 3D structures]] | |||
== References == | |||
<references/> | |||
__TOC__ | __TOC__ | ||
</StructureSection> | </StructureSection> | ||
[[Category: Large Structures]] | [[Category: Large Structures]] | ||
[[Category: Streptomyces griseoflavus]] | |||
[[Category: Gabdulkhakov A]] | [[Category: Gabdulkhakov A]] | ||
[[Category: Kolyadenko I]] | [[Category: Kolyadenko I]] | ||
[[Category: Tishchenko S]] | [[Category: Tishchenko S]] | ||