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| | ==Structure of Chitinase D from Serratia proteamaculans revealed an unusually constrained substrate binding site== | | ==Structure of Chitinase D from Serratia proteamaculans revealed an unusually constrained substrate binding site== |
| - | <StructureSection load='4lgx' size='340' side='right' caption='[[4lgx]], [[Resolution|resolution]] 1.49Å' scene=''> | + | <StructureSection load='4lgx' size='340' side='right'caption='[[4lgx]], [[Resolution|resolution]] 1.49Å' scene=''> |
| | == Structural highlights == | | == Structural highlights == |
| - | <table><tr><td colspan='2'>[[4lgx]] is a 1 chain structure with sequence from [http://en.wikipedia.org/wiki/Serp5 Serp5]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=4LGX OCA]. For a <b>guided tour on the structure components</b> use [http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=4LGX FirstGlance]. <br> | + | <table><tr><td colspan='2'>[[4lgx]] is a 1 chain structure with sequence from [https://en.wikipedia.org/wiki/Serratia_proteamaculans_568 Serratia proteamaculans 568]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=4LGX OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=4LGX FirstGlance]. <br> |
| - | </td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat"><scene name='pdbligand=ACT:ACETATE+ION'>ACT</scene>, <scene name='pdbligand=GOL:GLYCEROL'>GOL</scene></td></tr> | + | </td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=ACT:ACETATE+ION'>ACT</scene>, <scene name='pdbligand=GOL:GLYCEROL'>GOL</scene>, <scene name='pdbligand=OMT:S-DIOXYMETHIONINE'>OMT</scene></td></tr> |
| - | <tr id='NonStdRes'><td class="sblockLbl"><b>[[Non-Standard_Residue|NonStd Res:]]</b></td><td class="sblockDat"><scene name='pdbligand=OMT:S-DIOXYMETHIONINE'>OMT</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=4lgx FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=4lgx OCA], [https://pdbe.org/4lgx PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=4lgx RCSB], [https://www.ebi.ac.uk/pdbsum/4lgx PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=4lgx ProSAT]</span></td></tr> |
| - | <tr id='related'><td class="sblockLbl"><b>[[Related_structure|Related:]]</b></td><td class="sblockDat">[[3qok|3qok]]</td></tr>
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| - | <tr id='gene'><td class="sblockLbl"><b>[[Gene|Gene:]]</b></td><td class="sblockDat">Spro_2725 ([http://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?mode=Info&srchmode=5&id=399741 SERP5])</td></tr>
| + | |
| - | <tr id='activity'><td class="sblockLbl"><b>Activity:</b></td><td class="sblockDat"><span class='plainlinks'>[http://en.wikipedia.org/wiki/Chitinase Chitinase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=3.2.1.14 3.2.1.14] </span></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=4lgx FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=4lgx OCA], [http://pdbe.org/4lgx PDBe], [http://www.rcsb.org/pdb/explore.do?structureId=4lgx RCSB], [http://www.ebi.ac.uk/pdbsum/4lgx PDBsum], [http://prosat.h-its.org/prosat/prosatexe?pdbcode=4lgx ProSAT]</span></td></tr> | + | |
| | </table> | | </table> |
| | + | == Function == |
| | + | [https://www.uniprot.org/uniprot/A8GFD6_SERP5 A8GFD6_SERP5] |
| | <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: Chitinase]] | + | [[Category: Large Structures]] |
| - | [[Category: Serp5]] | + | [[Category: Serratia proteamaculans 568]] |
| - | [[Category: Kaur, P]] | + | [[Category: Kaur P]] |
| - | [[Category: Kumar, S]] | + | [[Category: Kumar S]] |
| - | [[Category: Madhuprakash, J]] | + | [[Category: Madhuprakash J]] |
| - | [[Category: Podile, A R]] | + | [[Category: Podile AR]] |
| - | [[Category: Sharma, S]] | + | [[Category: Sharma S]] |
| - | [[Category: Singh, A]] | + | [[Category: Singh A]] |
| - | [[Category: Singh, T P]] | + | [[Category: Singh TP]] |
| - | [[Category: Sinha, M]] | + | [[Category: Sinha M]] |
| - | [[Category: Hydrolase]]
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| - | [[Category: Tim barrel]]
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| Structural highlights
Function
A8GFD6_SERP5
Publication Abstract from PubMed
Serratia proteamaculans chitinase-D (SpChiD) has a unique combination of hydrolytic and transglycosylation (TG) activities. The TG activity of SpChiD can be used for large-scale production of chito-oligosaccharides (CHOS). The multiple activities (hydrolytic and/or chitobiase activities and TG) of SpChiD appear to be strongly influenced by the substrate-binding cleft. Here, we report the unique property of SpChiD substrate-binding cleft, wherein, the residues Tyr28, Val35 and Thr36 control chitobiase activity and the residues Trp160 and Trp290 are crucial for TG activity. Mutants with reduced (V35G and T36G/F) or no (SpChiDDelta30-42 and Y28A) chitobiase activity produced higher amounts of the quantifiable even-chain TG product with degree of polymerization (DP)-6, indicating that the chitobiase and TG activities are inversely related. In addition to its unprecedented catalytic properties, unlike other chitinases, the single modular SpChiD showed dual unfolding transitions. Ligand-induced thermal stability studies with the catalytically inactive mutant of SpChiD (E153A) showed that the transition temperature increased upon binding of CHOS with DP2-6. Isothermal titration calorimetry experiments revealed the exceptionally high binding affinities for E153A to CHOS with DP2-6. These observations strongly support that the architecture of SpChiD substrate-binding cleft adopted to control chitobiase and TG activities, in addition to usual chitinase-mediated hydrolysis.
Inverse relationship between chitobiase and transglycosylation activities of chitinase-D from Serratia proteamaculans revealed by mutational and biophysical analyses.,Madhuprakash J, Bobbili KB, Moerschbacher BM, Singh TP, Swamy MJ, Podile AR Sci Rep. 2015 Oct 23;5:15657. doi: 10.1038/srep15657. PMID:26493546[1]
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.
References
- ↑ Madhuprakash J, Bobbili KB, Moerschbacher BM, Singh TP, Swamy MJ, Podile AR. Inverse relationship between chitobiase and transglycosylation activities of chitinase-D from Serratia proteamaculans revealed by mutational and biophysical analyses. Sci Rep. 2015 Oct 23;5:15657. doi: 10.1038/srep15657. PMID:26493546 doi:http://dx.doi.org/10.1038/srep15657
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