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| | ==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]], [[Resolution|resolution]] 2.15Å' scene=''> |
| | == Structural highlights == | | == Structural highlights == |
| - | <table><tr><td colspan='2'>[[3x17]] is a 2 chain structure with sequence from [http://en.wikipedia.org/wiki/Uncultivated_bacterium Uncultivated bacterium]. 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 [http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=3X17 FirstGlance]. <br> | + | <table><tr><td colspan='2'>[[3x17]] is a 2 chain structure with sequence from [https://en.wikipedia.org/wiki/Uncultured_bacterium Uncultured bacterium]. 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"><scene name='pdbligand=CA:CALCIUM+ION'>CA</scene>, <scene name='pdbligand=ZN:ZINC+ION'>ZN</scene></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]] 2.15Å</td></tr> |
| - | <tr id='activity'><td class="sblockLbl"><b>Activity:</b></td><td class="sblockDat"><span class='plainlinks'>[http://en.wikipedia.org/wiki/Cellulase Cellulase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=3.2.1.4 3.2.1.4] </span></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> |
| - | <tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=3x17 FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=3x17 OCA], [http://pdbe.org/3x17 PDBe], [http://www.rcsb.org/pdb/explore.do?structureId=3x17 RCSB], [http://www.ebi.ac.uk/pdbsum/3x17 PDBsum], [http://prosat.h-its.org/prosat/prosatexe?pdbcode=3x17 ProSAT]</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> |
| | + | == Function == |
| | + | [https://www.uniprot.org/uniprot/W8PF21_9BACT W8PF21_9BACT] |
| | <div style="background-color:#fffaf0;"> | | <div style="background-color:#fffaf0;"> |
| | == Publication Abstract from PubMed == | | == Publication Abstract from PubMed == |
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| | __TOC__ | | __TOC__ |
| | </StructureSection> | | </StructureSection> |
| - | [[Category: Cellulase]] | + | [[Category: Large Structures]] |
| - | [[Category: Uncultivated bacterium]] | + | [[Category: Uncultured bacterium]] |
| - | [[Category: Angkawidjaja, C]] | + | [[Category: Angkawidjaja C]] |
| - | [[Category: Kanaya, S]] | + | [[Category: Kanaya S]] |
| - | [[Category: Okano, H]] | + | [[Category: Okano H]] |
| - | [[Category: Carbohydrate/sugar binding]]
| + | |
| - | [[Category: Hydrolase]]
| + | |
| Structural highlights
Function
W8PF21_9BACT
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[1]
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.
References
- ↑ Okano H, Kanaya E, Ozaki M, Angkawidjaja C, Kanaya S. Structure, activity, and stability of metagenome-derived glycoside hydrolase family 9 endoglucanase with an N-terminal Ig-like domain. Protein Sci. 2014 Dec 26. doi: 10.1002/pro.2632. PMID:25545469 doi:http://dx.doi.org/10.1002/pro.2632
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