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| - | ==CRYSTAL STRUCTURE OF INULOSUCRASE FROM LACTOBACILLUS JOHNSONII NCC533 IN COMPLEX WITH SUCROSE== | + | |
| - | <StructureSection load='2yfs' size='340' side='right' caption='[[2yfs]], [[Resolution|resolution]] 2.60Å' scene=''> | + | ==Crystal structure of inulosucrase from Lactobacillus johnsonii NCC533 in complex with sucrose== |
| | + | <StructureSection load='2yfs' size='340' side='right'caption='[[2yfs]], [[Resolution|resolution]] 2.60Å' scene=''> |
| | == Structural highlights == | | == Structural highlights == |
| - | <table><tr><td colspan='2'>[[2yfs]] is a 1 chain structure with sequence from [http://en.wikipedia.org/wiki/Lactobacillus_johnsonii Lactobacillus johnsonii]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=2YFS OCA]. <br> | + | <table><tr><td colspan='2'>[[2yfs]] is a 1 chain structure with sequence from [https://en.wikipedia.org/wiki/Lactobacillus_johnsonii Lactobacillus johnsonii]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=2YFS OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=2YFS FirstGlance]. <br> |
| - | </td></tr><tr><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat"><scene name='pdbligand=CA:CALCIUM+ION'>CA</scene>, <scene name='pdbligand=SO4:SULFATE+ION'>SO4</scene>, <scene name='pdbligand=SUC:SUCROSE'>SUC</scene><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]] 2.6Å</td></tr> |
| - | <tr><td class="sblockLbl"><b>[[Related_structure|Related:]]</b></td><td class="sblockDat">[[2yft|2yft]], [[2yfr|2yfr]]</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=FRU:FRUCTOSE'>FRU</scene>, <scene name='pdbligand=GLC:ALPHA-D-GLUCOSE'>GLC</scene>, <scene name='pdbligand=PRD_900003:sucrose'>PRD_900003</scene>, <scene name='pdbligand=SO4:SULFATE+ION'>SO4</scene></td></tr> |
| - | <tr><td class="sblockLbl"><b>Activity:</b></td><td class="sblockDat"><span class='plainlinks'>[http://en.wikipedia.org/wiki/Glucokinase Glucokinase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=2.7.1.2 2.7.1.2] </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=2yfs FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=2yfs OCA], [https://pdbe.org/2yfs PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=2yfs RCSB], [https://www.ebi.ac.uk/pdbsum/2yfs PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=2yfs ProSAT]</span></td></tr> |
| - | <tr><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=2yfs FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=2yfs OCA], [http://www.rcsb.org/pdb/explore.do?structureId=2yfs RCSB], [http://www.ebi.ac.uk/pdbsum/2yfs PDBsum]</span></td></tr> | + | </table> |
| - | <table> | + | == Function == |
| | + | [https://www.uniprot.org/uniprot/INUS_LACJO INUS_LACJO] Fructosyltransferase that catalyzes the polymerization of the fructose moiety of sucrose to produce inulin polymer and inulin oligosaccharides such as 1-kestose and nystose.<ref>PMID:18408060</ref> <ref>PMID:21801732</ref> |
| | <div style="background-color:#fffaf0;"> | | <div style="background-color:#fffaf0;"> |
| | == Publication Abstract from PubMed == | | == Publication Abstract from PubMed == |
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| | Crystal Structure of Inulosucrase from Lactobacillus: Insights into the Substrate Specificity and Product Specificity of GH68 Fructansucrases.,Pijning T, Anwar MA, Boger M, Dobruchowska JM, Leemhuis H, Kralj S, Dijkhuizen L, Dijkstra BW J Mol Biol. 2011 Sep 9;412(1):80-93. Epub 2011 Jul 23. PMID:21801732<ref>PMID:21801732</ref> | | Crystal Structure of Inulosucrase from Lactobacillus: Insights into the Substrate Specificity and Product Specificity of GH68 Fructansucrases.,Pijning T, Anwar MA, Boger M, Dobruchowska JM, Leemhuis H, Kralj S, Dijkhuizen L, Dijkstra BW J Mol Biol. 2011 Sep 9;412(1):80-93. Epub 2011 Jul 23. PMID:21801732<ref>PMID:21801732</ref> |
| | | | |
| - | From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br> | + | From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br> |
| | </div> | | </div> |
| | + | <div class="pdbe-citations 2yfs" style="background-color:#fffaf0;"></div> |
| | == References == | | == References == |
| | <references/> | | <references/> |
| | __TOC__ | | __TOC__ |
| | </StructureSection> | | </StructureSection> |
| - | [[Category: Inulosucrase]] | |
| | [[Category: Lactobacillus johnsonii]] | | [[Category: Lactobacillus johnsonii]] |
| - | [[Category: Anwar, M A.]] | + | [[Category: Large Structures]] |
| - | [[Category: Dijkhuizen, L.]] | + | [[Category: Anwar MA]] |
| - | [[Category: Dijkstra, B W.]] | + | [[Category: Dijkhuizen L]] |
| - | [[Category: Kralj, S.]] | + | [[Category: Dijkstra BW]] |
| - | [[Category: Leemhuis, H.]] | + | [[Category: Kralj S]] |
| - | [[Category: Pijning, T.]] | + | [[Category: Leemhuis H]] |
| - | [[Category: Fructosyltransferase]]
| + | [[Category: Pijning T]] |
| - | [[Category: Glycoside hydrolase family gh68]]
| + | |
| - | [[Category: Sugar utilization]]
| + | |
| - | [[Category: Transferase]]
| + | |
| Structural highlights
2yfs is a 1 chain structure with sequence from Lactobacillus johnsonii. Full crystallographic information is available from OCA. For a guided tour on the structure components use FirstGlance.
| | Method: | X-ray diffraction, Resolution 2.6Å |
| Ligands: | , , , , |
| Resources: | FirstGlance, OCA, PDBe, RCSB, PDBsum, ProSAT |
Function
INUS_LACJO Fructosyltransferase that catalyzes the polymerization of the fructose moiety of sucrose to produce inulin polymer and inulin oligosaccharides such as 1-kestose and nystose.[1] [2]
Publication Abstract from PubMed
Fructansucrases (FSs) catalyze a transfructosylation reaction with sucrose as substrate to produce fructo-oligosaccharides and fructan polymers that contain either beta-2,1 glycosidic linkages (inulin) or beta-2,6 linkages (levan). Levan-synthesizing FSs (levansucrases) have been most extensively investigated, while detailed information on inulosucrases is limited. Importantly, the molecular basis of the different product specificities of levansucrases and inulosucrases is poorly understood. We have elucidated the three-dimensional structure of a truncated active bacterial GH68 inulosucrase, InuJ of Lactobacillus johnsonii NCC533 (residues 145-708), in its apo form, with a bound substrate (sucrose), and with a transfructosylation product. The sucrose binding pocket and the sucrose binding mode are virtually identical with those of GH68 levansucrases, confirming that both enzyme types use the same fully conserved structural framework for the binding and cleavage of the donor substrate sucrose in the active site. The binding mode of the first transfructosylation product 1-kestose (Fru-beta(2-1)-Fru-alpha(2-1)-Glc, where Fru=fructose and Glc=glucose) in subsites -1 to +2 shows for the first time how inulin-type fructo-oligosaccharide bind in GH68 FS and how an inulin-type linkage can be formed. Surprisingly, observed interactions with the sugar in subsites +1 and +2 are provided by residues that are also present in levansucrases. The binding mode of 1-kestose and the presence of a more distant sucrose binding site suggest that residues beyond the +2 subsite, in particular residues from the nonconserved 1B-1C loop, determine product linkage type specificity in GH68 FSs.
Crystal Structure of Inulosucrase from Lactobacillus: Insights into the Substrate Specificity and Product Specificity of GH68 Fructansucrases.,Pijning T, Anwar MA, Boger M, Dobruchowska JM, Leemhuis H, Kralj S, Dijkhuizen L, Dijkstra BW J Mol Biol. 2011 Sep 9;412(1):80-93. Epub 2011 Jul 23. PMID:21801732[3]
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.
References
- ↑ Anwar MA, Kralj S, van der Maarel MJ, Dijkhuizen L. The probiotic Lactobacillus johnsonii NCC 533 produces high-molecular-mass inulin from sucrose by using an inulosucrase enzyme. Appl Environ Microbiol. 2008 Jun;74(11):3426-33. PMID:18408060 doi:10.1128/AEM.00377-08
- ↑ Pijning T, Anwar MA, Boger M, Dobruchowska JM, Leemhuis H, Kralj S, Dijkhuizen L, Dijkstra BW. Crystal Structure of Inulosucrase from Lactobacillus: Insights into the Substrate Specificity and Product Specificity of GH68 Fructansucrases. J Mol Biol. 2011 Sep 9;412(1):80-93. Epub 2011 Jul 23. PMID:21801732 doi:10.1016/j.jmb.2011.07.031
- ↑ Pijning T, Anwar MA, Boger M, Dobruchowska JM, Leemhuis H, Kralj S, Dijkhuizen L, Dijkstra BW. Crystal Structure of Inulosucrase from Lactobacillus: Insights into the Substrate Specificity and Product Specificity of GH68 Fructansucrases. J Mol Biol. 2011 Sep 9;412(1):80-93. Epub 2011 Jul 23. PMID:21801732 doi:10.1016/j.jmb.2011.07.031
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