4flo

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==Crystal structure of Amylosucrase double mutant A289P-F290C from Neisseria polysaccharea==
==Crystal structure of Amylosucrase double mutant A289P-F290C from Neisseria polysaccharea==
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<StructureSection load='4flo' size='340' side='right' caption='[[4flo]], [[Resolution|resolution]] 2.20&Aring;' scene=''>
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<StructureSection load='4flo' size='340' side='right'caption='[[4flo]], [[Resolution|resolution]] 2.20&Aring;' scene=''>
== Structural highlights ==
== Structural highlights ==
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<table><tr><td colspan='2'>[[4flo]] is a 1 chain structure with sequence from [http://en.wikipedia.org/wiki/Neisseria_polysaccharea Neisseria polysaccharea]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=4FLO OCA]. For a <b>guided tour on the structure components</b> use [http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=4FLO FirstGlance]. <br>
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<table><tr><td colspan='2'>[[4flo]] is a 1 chain structure with sequence from [https://en.wikipedia.org/wiki/Neisseria_polysaccharea Neisseria polysaccharea]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=4FLO OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=4FLO FirstGlance]. <br>
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</td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat"><scene name='pdbligand=GOL:GLYCEROL'>GOL</scene>, <scene name='pdbligand=P6G:HEXAETHYLENE+GLYCOL'>P6G</scene>, <scene name='pdbligand=TRS:2-AMINO-2-HYDROXYMETHYL-PROPANE-1,3-DIOL'>TRS</scene></td></tr>
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</td></tr><tr id='method'><td class="sblockLbl"><b>[[Empirical_models|Method:]]</b></td><td class="sblockDat" id="methodDat">X-ray diffraction, [[Resolution|Resolution]] 2.2&#8491;</td></tr>
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<tr id='related'><td class="sblockLbl"><b>[[Related_structure|Related:]]</b></td><td class="sblockDat">[[1g5a|1g5a]], [[4flq|4flq]], [[4flr|4flr]], [[4fls|4fls]]</td></tr>
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<tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=GOL:GLYCEROL'>GOL</scene>, <scene name='pdbligand=P6G:HEXAETHYLENE+GLYCOL'>P6G</scene>, <scene name='pdbligand=TRS:2-AMINO-2-HYDROXYMETHYL-PROPANE-1,3-DIOL'>TRS</scene></td></tr>
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<tr id='gene'><td class="sblockLbl"><b>[[Gene|Gene:]]</b></td><td class="sblockDat">ams ([http://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?mode=Info&srchmode=5&id=489 Neisseria polysaccharea])</td></tr>
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<tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[https://proteopedia.org/fgij/fg.htm?mol=4flo FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=4flo OCA], [https://pdbe.org/4flo PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=4flo RCSB], [https://www.ebi.ac.uk/pdbsum/4flo PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=4flo ProSAT]</span></td></tr>
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<tr id='activity'><td class="sblockLbl"><b>Activity:</b></td><td class="sblockDat"><span class='plainlinks'>[http://en.wikipedia.org/wiki/Amylosucrase Amylosucrase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=2.4.1.4 2.4.1.4] </span></td></tr>
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<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=4flo FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=4flo OCA], [http://www.rcsb.org/pdb/explore.do?structureId=4flo RCSB], [http://www.ebi.ac.uk/pdbsum/4flo PDBsum]</span></td></tr>
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</table>
</table>
== Function ==
== Function ==
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[[http://www.uniprot.org/uniprot/AMYS_NEIPO AMYS_NEIPO]] Catalyzes the synthesis of alpha-glucan from sucrose. Catalyzes, in addition, sucrose hydrolysis, maltose and maltotriose synthesis by successive transfers of the glucosyl moiety of sucrose onto the released glucose, and finally turanose and trehalulose synthesis, these two sucrose isomers being obtained by glucosyl transfer onto fructose.<ref>PMID:9882648</ref>
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[https://www.uniprot.org/uniprot/AMYS_NEIPO AMYS_NEIPO] Catalyzes the synthesis of alpha-glucan from sucrose. Catalyzes, in addition, sucrose hydrolysis, maltose and maltotriose synthesis by successive transfers of the glucosyl moiety of sucrose onto the released glucose, and finally turanose and trehalulose synthesis, these two sucrose isomers being obtained by glucosyl transfer onto fructose.<ref>PMID:9882648</ref>
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<div style="background-color:#fffaf0;">
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== Publication Abstract from PubMed ==
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Iterative saturation mutagenesis and combinatorial active site saturation focused on vicinal amino acids were used to alter the acceptor specificity of amylosucrase from Neisseria polysaccharea , a sucrose-utilizing alpha-transglucosidase, and sort out improved variants. From the screening of three semirational sublibraries accounting in total for 20 000 variants, we report here the isolation of three double mutants of N. polysaccharea amylosucrase displaying a spectacular specificity enhancement toward both sucrose, the donor substrate, and the allyl 2-acetamido-2-deoxy-alpha-d-glucopyranoside acceptor as compared to the wild-type enzyme. Such levels of activity improvement have never been reported before for this class of carbohydrate-active enzymes. X-ray structure of the best performing enzymes supported by molecular dynamics simulations showed local rigidity of the -1 subsite as well as flexibility of loops involved in active site topology, which both account for the enhanced catalytic performances of the mutants. The study well illustrates the importance of taking into account the local conformation of catalytic residues as well as protein dynamics during the catalytic process, when designing enzyme libraries.
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Applying pairwise combinations of amino Acid mutations for sorting out highly efficient glucosylation tools for chemo-enzymatic synthesis of bacterial oligosaccharides.,Champion E, Guerin F, Moulis C, Barbe S, Tran TH, Morel S, Descroix K, Monsan P, Mourey L, Mulard LA, Tranier S, Remaud-Simeon M, Andre I J Am Chem Soc. 2012 Nov 14;134(45):18677-88. doi: 10.1021/ja306845b. Epub 2012, Oct 30. PMID:23072374<ref>PMID:23072374</ref>
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From MEDLINE&reg;/PubMed&reg;, a database of the U.S. National Library of Medicine.<br>
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</div>
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== References ==
== References ==
<references/>
<references/>
__TOC__
__TOC__
</StructureSection>
</StructureSection>
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[[Category: Amylosucrase]]
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[[Category: Large Structures]]
[[Category: Neisseria polysaccharea]]
[[Category: Neisseria polysaccharea]]
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[[Category: Andre, I]]
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[[Category: Andre I]]
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[[Category: Barbe, S]]
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[[Category: Barbe S]]
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[[Category: Champion, E]]
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[[Category: Champion E]]
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[[Category: Descroix, K]]
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[[Category: Descroix K]]
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[[Category: Guerin, F]]
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[[Category: Guerin F]]
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[[Category: Monsan, P]]
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[[Category: Monsan P]]
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[[Category: Morel, S]]
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[[Category: Morel S]]
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[[Category: Moulis, C]]
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[[Category: Moulis C]]
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[[Category: Mourey, L]]
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[[Category: Mourey L]]
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[[Category: Mulard, L A]]
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[[Category: Mulard LA]]
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[[Category: Remaud-Simeon, M]]
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[[Category: Remaud-Simeon M]]
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[[Category: Tran, T H]]
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[[Category: Tran TH]]
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[[Category: Tranier, S]]
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[[Category: Tranier S]]
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[[Category: Amylose synthesis]]
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[[Category: Beta/alpha-barrel]]
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[[Category: Carbohydrate]]
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[[Category: Glucosyltransferase]]
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[[Category: Glycoside hydrolase]]
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[[Category: Sucrose isomerization]]
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[[Category: Transferase]]
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Current revision

Crystal structure of Amylosucrase double mutant A289P-F290C from Neisseria polysaccharea

PDB ID 4flo

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