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| | ==Crystal Structure of Pasteurella multocida N-Acetyl-D-Neuraminic acid lyase K164 mutant complexed with N-Glycolylneuraminic acid== | | ==Crystal Structure of Pasteurella multocida N-Acetyl-D-Neuraminic acid lyase K164 mutant complexed with N-Glycolylneuraminic acid== |
| - | <StructureSection load='4img' size='340' side='right' caption='[[4img]], [[Resolution|resolution]] 1.85Å' scene=''> | + | <StructureSection load='4img' size='340' side='right'caption='[[4img]], [[Resolution|resolution]] 1.85Å' scene=''> |
| | == Structural highlights == | | == Structural highlights == |
| - | <table><tr><td colspan='2'>[[4img]] is a 2 chain structure with sequence from [http://en.wikipedia.org/wiki/Pasteurella_multocida_subsp._gallicida_p1059 Pasteurella multocida subsp. gallicida p1059]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=4IMG OCA]. For a <b>guided tour on the structure components</b> use [http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=4IMG FirstGlance]. <br> | + | <table><tr><td colspan='2'>[[4img]] is a 2 chain structure with sequence from [https://en.wikipedia.org/wiki/Pasteurella_multocida_subsp._gallicida_P1059 Pasteurella multocida subsp. gallicida P1059]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=4IMG OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=4IMG FirstGlance]. <br> |
| - | </td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat"><scene name='pdbligand=ME2:1-ETHOXY-2-(2-METHOXYETHOXY)ETHANE'>ME2</scene>, <scene name='pdbligand=NGF:3,5-DIDEOXY-5-[(HYDROXYACETYL)AMINO]-D-GLYCERO-D-GALACTO-NON-2-ULOSONIC+ACID'>NGF</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]] 1.85Å</td></tr> |
| - | <tr id='related'><td class="sblockLbl"><b>[[Related_structure|Related:]]</b></td><td class="sblockDat">[[4imc|4imc]], [[4imd|4imd]], [[4ime|4ime]], [[4imf|4imf]]</td></tr>
| + | <tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=ME2:1-ETHOXY-2-(2-METHOXYETHOXY)ETHANE'>ME2</scene>, <scene name='pdbligand=NGF:3,5-DIDEOXY-5-[(HYDROXYACETYL)AMINO]-D-GLYCERO-D-GALACTO-NON-2-ULOSONIC+ACID'>NGF</scene></td></tr> |
| - | <tr id='gene'><td class="sblockLbl"><b>[[Gene|Gene:]]</b></td><td class="sblockDat">nanA, PM1715 ([http://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?mode=Info&srchmode=5&id=1169409 Pasteurella multocida subsp. gallicida P1059])</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=4img FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=4img OCA], [https://pdbe.org/4img PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=4img RCSB], [https://www.ebi.ac.uk/pdbsum/4img PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=4img ProSAT]</span></td></tr> |
| - | <tr id='activity'><td class="sblockLbl"><b>Activity:</b></td><td class="sblockDat"><span class='plainlinks'>[http://en.wikipedia.org/wiki/N-acetylneuraminate_lyase N-acetylneuraminate lyase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=4.1.3.3 4.1.3.3] </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=4img FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=4img OCA], [http://pdbe.org/4img PDBe], [http://www.rcsb.org/pdb/explore.do?structureId=4img RCSB], [http://www.ebi.ac.uk/pdbsum/4img PDBsum], [http://prosat.h-its.org/prosat/prosatexe?pdbcode=4img ProSAT]</span></td></tr> | + | |
| | </table> | | </table> |
| | == Function == | | == Function == |
| - | [[http://www.uniprot.org/uniprot/NANA_PASMU NANA_PASMU]] Catalyzes the cleavage of N-acetylneuraminic acid (sialic acid) to form pyruvate and N-acetylmannosamine via a Schiff base intermediate (By similarity). | + | [https://www.uniprot.org/uniprot/NANA_PASMU NANA_PASMU] Catalyzes the cleavage of N-acetylneuraminic acid (sialic acid) to form pyruvate and N-acetylmannosamine via a Schiff base intermediate (By similarity). |
| - | <div style="background-color:#fffaf0;">
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| - | == Publication Abstract from PubMed ==
| + | |
| - | <i>N</i>-Acetylneuraminate lyases (NALs) or sialic acid aldolases catalyze the reversible aldol cleavage of <i>N</i>-acetylneuraminic acid (Neu5Ac, the most common form of sialic acid) to form pyruvate and <i>N</i>-acetyl-D-mannosamine (ManNAc). Although equilibrium favors sialic acid cleavage, these enzymes can be used for high-yield chemoenzymatic synthesis of structurally diverse sialic acids in the presence of excess pyruvate. Engineering these enzymes to synthesize structurally modified natural sialic acids and their non-natural derivatives holds promise in creating novel therapeutic agents. Atomic resolution structures of these enzymes will greatly assist in guiding mutagenic and modeling studies to engineer enzymes with altered substrate specificity. We report here the crystal structures of wild-type <i>Pasteurella multocida N</I>-acetylneuraminate lyase and its K164A mutant. Like other bacterial lyases, it assembles into a homotetramer with each monomer folding into a classic (beta/alpha)<sub>8</sub> TIM barrel. Two wild-type structures were determined; in the absence of substrates, and trapped in a Schiff base intermediate between Lys164 and pyruvate, respectively. Three structures of the K164A variant were determined: one in the absence of substrates and two binary complexes with <i>N</i>-acetylneuraminic acid (Neu5Ac) and <i>N</i>-glycolylneuraminic acid (Neu5Gc), respectively. Both sialic acids bind to the active site in the open-chain ketone form of the monosaccharide. The structures reveal that every hydroxyl group of the linear sugars makes hydrogen bond interactions with the enzyme and the residues that determine specificity were identified. Additionally, the structures lend some clues in explaining the natural discrimination of sialic acid substrates between the <i>P. multocida</i> and <i>E. coli</i> NALs.
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| - | Structural basis for substrate specificity and mechanism of <i>N</i>-acetyl-D-neuraminic acid lyase from <i>Pasteurella multocida.</i>,Huynh N, Aye A, Li Y, Yu H, Cao H, Tiwari VK, Shin DW, Chen X, Fisher AJ Biochemistry. 2013 Oct 23. PMID:24152047<ref>PMID:24152047</ref>
| + | ==See Also== |
| - | | + | *[[N-acetylneuraminate lyase 3D structures|N-acetylneuraminate lyase 3D structures]] |
| - | From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br>
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| - | </div>
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| - | <div class="pdbe-citations 4img" style="background-color:#fffaf0;"></div>
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| - | == References ==
| + | |
| - | <references/>
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| | __TOC__ | | __TOC__ |
| | </StructureSection> | | </StructureSection> |
| - | [[Category: N-acetylneuraminate lyase]] | + | [[Category: Large Structures]] |
| - | [[Category: Pasteurella multocida subsp. gallicida p1059]] | + | [[Category: Pasteurella multocida subsp. gallicida P1059]] |
| - | [[Category: Fisher, A J]] | + | [[Category: Fisher AJ]] |
| - | [[Category: Huynh, N]] | + | [[Category: Huynh N]] |
| - | [[Category: Lyase]]
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| - | [[Category: Schiff base]]
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| - | [[Category: Tim barrel]]
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