3d6e
From Proteopedia
(Difference between revisions)
(New page: '''Unreleased structure''' The entry 3d6e is ON HOLD until Paper Publication Authors: Fita, I., Planas, A., Calisto, B.M., Addington, T. Description: Crystal structure of the engineere...) |
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| - | '''Unreleased structure''' | ||
| - | + | ==Crystal structure of the engineered 1,3-1,4-beta-glucanase protein from Bacillus licheniformis== | |
| + | <StructureSection load='3d6e' size='340' side='right'caption='[[3d6e]], [[Resolution|resolution]] 2.40Å' scene=''> | ||
| + | == Structural highlights == | ||
| + | <table><tr><td colspan='2'>[[3d6e]] is a 2 chain structure with sequence from [https://en.wikipedia.org/wiki/Bacillus_licheniformis Bacillus licheniformis]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=3D6E OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=3D6E 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]] 2.4Å</td></tr> | ||
| + | <tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=CA:CALCIUM+ION'>CA</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=3d6e FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=3d6e OCA], [https://pdbe.org/3d6e PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=3d6e RCSB], [https://www.ebi.ac.uk/pdbsum/3d6e PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=3d6e ProSAT]</span></td></tr> | ||
| + | </table> | ||
| + | == Function == | ||
| + | [https://www.uniprot.org/uniprot/GUB_BACLI GUB_BACLI] | ||
| + | == Evolutionary Conservation == | ||
| + | [[Image:Consurf_key_small.gif|200px|right]] | ||
| + | Check<jmol> | ||
| + | <jmolCheckbox> | ||
| + | <scriptWhenChecked>; select protein; define ~consurf_to_do selected; consurf_initial_scene = true; script "/wiki/ConSurf/d6/3d6e_consurf.spt"</scriptWhenChecked> | ||
| + | <scriptWhenUnchecked>script /wiki/extensions/Proteopedia/spt/initialview01.spt</scriptWhenUnchecked> | ||
| + | <text>to colour the structure by Evolutionary Conservation</text> | ||
| + | </jmolCheckbox> | ||
| + | </jmol>, as determined by [http://consurfdb.tau.ac.il/ ConSurfDB]. You may read the [[Conservation%2C_Evolutionary|explanation]] of the method and the full data available from [http://bental.tau.ac.il/new_ConSurfDB/main_output.php?pdb_ID=3d6e ConSurf]. | ||
| + | <div style="clear:both"></div> | ||
| + | <div style="background-color:#fffaf0;"> | ||
| + | == Publication Abstract from PubMed == | ||
| + | Family 16 carbohydrate active enzyme members Bacillus licheniformis 1,3-1,4-beta-glucanase and Populus tremula x tremuloides xyloglucan endotransglycosylase (XET16-34) are highly structurally related but display different substrate specificities. Although the first binds linear gluco-oligosaccharides, the second binds branched xylogluco-oligosaccharides. Prior engineered nucleophile mutants of both enzymes are glycosynthases that catalyze the condensation between a glycosyl fluoride donor and a glycoside acceptor. With the aim of expanding the glycosynthase technology to produce designer oligosaccharides consisting of hybrids between branched xylogluco- and linear gluco-oligosaccharides, enzyme engineering on the negative subsites of 1,3-1,4-beta-glucanase to accept branched substrates has been undertaken. Removal of the 1,3-1,4-beta-glucanase major loop and replacement with that of XET16-34 to open the binding cleft resulted in a folded protein, which still maintained some beta-glucan hydrolase activity, but the corresponding nucleophile mutant did not display glycosynthase activity with either linear or branched glycosyl donors. Next, point mutations of the 1,3-1,4-beta-glucanase beta-sheets forming the binding site cleft were mutated to resemble XET16-34 residues. The final chimeric protein acquired binding affinity for xyloglucan and did not bind beta-glucan. Therefore, binding specificity has been re-engineered, but affinity was low and the nucleophile mutant of the chimeric enzyme did not show glycosynthase activity to produce the target hybrid oligosaccharides. Structural analysis by X-ray crystallography explains these results in terms of changes in the protein structure and highlights further engineering approaches toward introducing the desired activity. Proteins 2010. (c) 2010 Wiley-Liss, Inc. | ||
| - | + | Re-engineering specificity in 1,3-1, 4-beta-glucanase to accept branched xyloglucan substrates.,Addington T, Calisto B, Alfonso-Prieto M, Rovira C, Fita I, Planas A Proteins. 2010 Sep 22. PMID:21069723<ref>PMID:21069723</ref> | |
| - | + | From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br> | |
| + | </div> | ||
| + | <div class="pdbe-citations 3d6e" style="background-color:#fffaf0;"></div> | ||
| - | + | ==See Also== | |
| - | + | *[[Glucanase 3D structures|Glucanase 3D structures]] | |
| + | == References == | ||
| + | <references/> | ||
| + | __TOC__ | ||
| + | </StructureSection> | ||
| + | [[Category: Bacillus licheniformis]] | ||
| + | [[Category: Large Structures]] | ||
| + | [[Category: Addington T]] | ||
| + | [[Category: Calisto BM]] | ||
| + | [[Category: Fita I]] | ||
| + | [[Category: Planas A]] | ||
Current revision
Crystal structure of the engineered 1,3-1,4-beta-glucanase protein from Bacillus licheniformis
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