|
|
| Line 1: |
Line 1: |
|
| |
|
| ==Crystal structure of metagenome-derived glycoside hydrolase family 9 endoglucanase== | | ==Crystal structure of metagenome-derived glycoside hydrolase family 9 endoglucanase== |
| <StructureSection load='3x17' size='340' side='right'caption='[[3x17]], [[Resolution|resolution]] 2.15Å' scene=''> | | <StructureSection load='3x17' size='340' side='right'caption='[[3x17]]' scene=''> |
| == Structural highlights == | | == Structural highlights == |
| <table><tr><td colspan='2'>[[3x17]] is a 2 chain structure. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=3X17 OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=3X17 FirstGlance]. <br> | | <table><tr><td colspan='2'>Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=3X17 OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=3X17 FirstGlance]. <br> |
| </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>, <scene name='pdbligand=ZN:ZINC+ION'>ZN</scene></td></tr> | | </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=3x17 FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=3x17 OCA], [https://pdbe.org/3x17 PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=3x17 RCSB], [https://www.ebi.ac.uk/pdbsum/3x17 PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=3x17 ProSAT]</span></td></tr> |
| <tr id='activity'><td class="sblockLbl"><b>Activity:</b></td><td class="sblockDat"><span class='plainlinks'>[https://en.wikipedia.org/wiki/Cellulase Cellulase], with EC number [https://www.brenda-enzymes.info/php/result_flat.php4?ecno=3.2.1.4 3.2.1.4] </span></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=3x17 FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=3x17 OCA], [https://pdbe.org/3x17 PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=3x17 RCSB], [https://www.ebi.ac.uk/pdbsum/3x17 PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=3x17 ProSAT]</span></td></tr> | |
| </table> | | </table> |
| <div style="background-color:#fffaf0;">
| |
| == Publication Abstract from PubMed ==
| |
| A metagenome-derived glycoside hydrolase family 9 enzyme with an N-terminal immunoglobulin-like (Ig-like) domain, leaf-branch compost (LC)-CelG, was characterized and its crystal structure was determined. LC-CelG did not hydrolyze p-nitrophenyl cellobioside but hydrolyzed CM-cellulose, indicating that it is endoglucanase. LC-CelG exhibited the highest activity at 70 degrees C and >80% of the maximal activity at a broad pH range of 5-9. Its denaturation temperature was 81.4 degrees C, indicating that LC-CelG is a thermostable enzyme. The structure of LC-CelG resembles those of CelD from Clostridium thermocellum (CtCelD), Cel9A from Alicyclobacillus acidocaldarius (AaCel9A), and cellobiohydrolase CbhA from C. thermocellum (CtCbhA), which show relatively low (29-31%) amino acid sequence identities to LC-CelG. Three acidic active site residues are conserved as Asp194, Asp197, and Glu558 in LC-CelG. Ten of the thirteen residues that form the substrate binding pocket of AaCel9A are conserved in LC-CelG. Removal of the Ig-like domain reduced the activity and stability of LC-CelG by 100-fold and 6.3 degrees C, respectively. Removal of the Gln40- and Asp99-mediated interactions between the Ig-like and catalytic domains destabilized LC-CelG by 5.0 degrees C without significantly affecting its activity. These results suggest that the Ig-like domain contributes to the stabilization of LC-CelG mainly due to the Gln40- and Asp99-mediated interactions. Because the LC-CelG derivative lacking the Ig-like domain accumulated in Escherichia coli cells mostly in an insoluble form and this derivative accumulated in a soluble form exhibited very weak activity, the Ig-like domain may be required to make the conformation of the active site functional and prevent aggregation of the catalytic domain.
| |
|
| |
| Structure, activity, and stability of metagenome-derived glycoside hydrolase family 9 endoglucanase with an N-terminal Ig-like domain.,Okano H, Kanaya E, Ozaki M, Angkawidjaja C, Kanaya S Protein Sci. 2014 Dec 26. doi: 10.1002/pro.2632. PMID:25545469<ref>PMID:25545469</ref>
| |
|
| |
| From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br>
| |
| </div>
| |
| <div class="pdbe-citations 3x17" style="background-color:#fffaf0;"></div>
| |
| == References ==
| |
| <references/>
| |
| __TOC__ | | __TOC__ |
| </StructureSection> | | </StructureSection> |
| [[Category: Cellulase]]
| |
| [[Category: Large Structures]] | | [[Category: Large Structures]] |
| [[Category: Angkawidjaja, C]] | | [[Category: Angkawidjaja C]] |
| [[Category: Kanaya, S]] | | [[Category: Kanaya S]] |
| [[Category: Okano, H]] | | [[Category: Okano H]] |
| [[Category: Carbohydrate/sugar binding]]
| |
| [[Category: Hydrolase]]
| |
