View source for 7vsx


==Crystal structure of QL-nanoKAZ (Reverse mutant of nanoKAZ with L18Q and V27L)==
<StructureSection load='7vsx' size='340' side='right'caption='[[7vsx]], [[Resolution|resolution]] 1.70&Aring;' scene=''>
== Structural highlights ==
<table><tr><td colspan='2'>[[7vsx]] is a 1 chain structure with sequence from [https://en.wikipedia.org/wiki/Oplophorus_gracilirostris Oplophorus gracilirostris]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=7VSX OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=7VSX 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]] 1.698&#8491;</td></tr>
<tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=MES:2-(N-MORPHOLINO)-ETHANESULFONIC+ACID'>MES</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=7vsx FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=7vsx OCA], [https://pdbe.org/7vsx PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=7vsx RCSB], [https://www.ebi.ac.uk/pdbsum/7vsx PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=7vsx ProSAT]</span></td></tr>
</table>
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== Publication Abstract from PubMed ==
Native Oplophorus luciferase (OpLase) and its catalytic 19 kDa protein (wild KAZ) show highest luminescence activity with coelenterazine (CTZ) among CTZ analogs. Mutated wild KAZ with 16 amino acid substitutions (nanoKAZ/nanoLuc) utilizes bis-coelenterazine (bis-CTZ) as the preferred substrate and exhibits over 10-fold higher maximum intensity than CTZ. To understand the substrate selectivity of nanoKAZ between CTZ and bis-CTZ, we prepared the reverse mutants of nanoKAZ by amino acid replacements with the original amino acid residue of wild KAZ. The reverse mutant with L18Q and V27L substitutions (QL-nanoKAZ) exhibited 2.6-fold higher maximum intensity with CTZ than that of nanoKAZ with bis-CTZ. The catalytic properties of QL-nanoKAZ including substrate specificity, luminescence spectrum, luminescence kinetics, luminescence products of CTZ, and luminescence inhibition by deaza-CTZ analogs were characterized and were compared with other CTZ-utilizing luciferases such as Gaussia and Renilla luciferases. Thus, QL-nanoKAZ with CTZ could be used as a potential reporter protein for various luminescence assay systems. Furthermore, the crystal structure of QL-nanoKAZ was determined at 1.70 A resolution. The reverse mutation at the L18Q and V27L positions of alpha2-helix in nanoKAZ led to changes in the local structures of the alpha4-helix and the beta6- and beta7-sheets, and might enhance its binding affinity and oxidation efficiency with CTZ to emit light.

Reverse mutants of the catalytic 19 kDa mutant protein (nanoKAZ/nanoLuc) from Oplophorus luciferase with coelenterazine as preferred substrate.,Inouye S, Sato JI, Sahara-Miura Y, Tomabechi Y, Sumida Y, Sekine SI, Shirouzu M, Hosoya T PLoS One. 2022 Sep 21;17(9):e0272992. doi: 10.1371/journal.pone.0272992., eCollection 2022. PMID:36129943<ref>PMID:36129943</ref>

From MEDLINE&reg;/PubMed&reg;, a database of the U.S. National Library of Medicine.<br>
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<div class="pdbe-citations 7vsx" style="background-color:#fffaf0;"></div>
== References ==
<references/>
__TOC__
</StructureSection>
[[Category: Large Structures]]
[[Category: Oplophorus gracilirostris]]
[[Category: Satoshi I]]
[[Category: Sekine S]]
[[Category: Shirouzu M]]
[[Category: Takamitsu H]]
[[Category: Tomabechi Y]]