9qez
From Proteopedia
(Difference between revisions)
(New page: '''Unreleased structure''' The entry 9qez is ON HOLD Authors: Mohsin, I., Papageorgiou, A.C. Description: Carbonic anhydrase mutant Category: Unreleased Structures [[Category: Papa...) |
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| - | '''Unreleased structure''' | ||
| - | + | ==Carbonic anhydrase mutant== | |
| + | <StructureSection load='9qez' size='340' side='right'caption='[[9qez]], [[Resolution|resolution]] 2.10Å' scene=''> | ||
| + | == Structural highlights == | ||
| + | <table><tr><td colspan='2'>[[9qez]] is a 10 chain structure with sequence from [https://en.wikipedia.org/wiki/Caloramator_australicus_RC3 Caloramator australicus RC3] and [https://en.wikipedia.org/wiki/Unidentified Unidentified]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=9QEZ OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=9QEZ 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]] 2.1Å</td></tr> | ||
| + | <tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=EPE:4-(2-HYDROXYETHYL)-1-PIPERAZINE+ETHANESULFONIC+ACID'>EPE</scene>, <scene name='pdbligand=MG:MAGNESIUM+ION'>MG</scene>, <scene name='pdbligand=NA:SODIUM+ION'>NA</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'>[https://proteopedia.org/fgij/fg.htm?mol=9qez FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=9qez OCA], [https://pdbe.org/9qez PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=9qez RCSB], [https://www.ebi.ac.uk/pdbsum/9qez PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=9qez ProSAT]</span></td></tr> | ||
| + | </table> | ||
| + | <div style="background-color:#fffaf0;"> | ||
| + | == Publication Abstract from PubMed == | ||
| + | The 16S microbial community profiling of a metagenomics library from geothermal spring at Lisvori (Lesvos island, Greece) enabled the identification of a putative sequence exhibiting 95% identity to the gamma-type carbonic anhydrase (gamma-CA) from Caloramator australicus (gamma-CaCA). The sequence of gamma-CaCA was amplified by PCR, cloned, and expressed in E. coli. Activity assays showed that gamma-CaCA possesses very low, but detectable, anhydrase activity, while exhibiting no measurable esterase activity. Differential scanning fluorimetry (DSF) revealed that the enzyme shows high thermal stability with a melting temperature (T(m)) approximately 65-75 degrees C in the pH range between 5.5 and 9.0. The structure of gamma-CaCA was determined by X-ray crystallography at 1.11 A resolution, the highest resolution reported so far for a gamma-CA. The enzyme was crystallized as a trimer in the crystallographic asymmetric unit and contains three zinc-binding sites, one at each interface of neighboring subunits of the trimer. Structure-based rational design enabled the design and creation of a mutant enzyme (gamma-CaCAmut) which possessed a heptapeptide insertion at the active-site loop and two-point mutations. Kinetic analysis demonstrated that gamma-CaCAmut was successfully converted into a catalytically active esterase indicating successful activity gain through structure-guided engineering. The thermostability of gamma-CaCAmut was significantly increased, aligning with the thermostability typically observed in hyperthermostable enzymes. X-ray crystallographic analysis of the gamma-CaCAmut structure at 2.1 A resolution, provided detailed structural insights into how the mutations impact the overall enzyme structure, function, and thermostability. These findings provide valuable structural and functional insights into gamma-CAs and demonstrate a strategy for converting an inactive enzyme into a catalytically active form through rational design. | ||
| - | + | Structural and functional characterization of a metagenomically derived gamma-type carbonic anhydrase and its engineering into a hyperthermostable esterase.,Bodourian CS, Imran M, Georgakis ND, Papageorgiou AC, Labrou NE Protein Sci. 2025 Dec;34(12):e70396. doi: 10.1002/pro.70396. PMID:41294346<ref>PMID:41294346</ref> | |
| - | + | From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br> | |
| - | [[Category: | + | </div> |
| - | [[Category: | + | <div class="pdbe-citations 9qez" style="background-color:#fffaf0;"></div> |
| - | [[Category: Mohsin | + | == References == |
| + | <references/> | ||
| + | __TOC__ | ||
| + | </StructureSection> | ||
| + | [[Category: Caloramator australicus RC3]] | ||
| + | [[Category: Large Structures]] | ||
| + | [[Category: Unidentified]] | ||
| + | [[Category: Mohsin I]] | ||
| + | [[Category: Papageorgiou AC]] | ||
Current revision
Carbonic anhydrase mutant
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