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| | ==Crystal structure of a 4-oxalocrotonate tautomerase mutant in complex with nitrostyrene at 2.3 Angstrom== | | ==Crystal structure of a 4-oxalocrotonate tautomerase mutant in complex with nitrostyrene at 2.3 Angstrom== |
| - | <StructureSection load='5clo' size='340' side='right' caption='[[5clo]], [[Resolution|resolution]] 2.30Å' scene=''> | + | <StructureSection load='5clo' size='340' side='right'caption='[[5clo]], [[Resolution|resolution]] 2.30Å' scene=''> |
| | == Structural highlights == | | == Structural highlights == |
| - | <table><tr><td colspan='2'>[[5clo]] is a 18 chain structure. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=5CLO OCA]. For a <b>guided tour on the structure components</b> use [http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=5CLO FirstGlance]. <br> | + | <table><tr><td colspan='2'>[[5clo]] is a 18 chain structure with sequence from [https://en.wikipedia.org/wiki/Pseudomonas_putida Pseudomonas putida]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=5CLO OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=5CLO FirstGlance]. <br> |
| - | </td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat"><scene name='pdbligand=NS8:TRANS+BETA+NITROSTYRENE'>NS8</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]] 2.3Å</td></tr> |
| - | <tr id='activity'><td class="sblockLbl"><b>Activity:</b></td><td class="sblockDat"><span class='plainlinks'>[http://en.wikipedia.org/wiki/2-hydroxymuconate_tautomerase 2-hydroxymuconate tautomerase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=5.3.2.6 5.3.2.6] </span></td></tr> | + | <tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=NS8:TRANS+BETA+NITROSTYRENE'>NS8</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=5clo FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=5clo OCA], [http://pdbe.org/5clo PDBe], [http://www.rcsb.org/pdb/explore.do?structureId=5clo RCSB], [http://www.ebi.ac.uk/pdbsum/5clo PDBsum]</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=5clo FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=5clo OCA], [https://pdbe.org/5clo PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=5clo RCSB], [https://www.ebi.ac.uk/pdbsum/5clo PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=5clo ProSAT]</span></td></tr> |
| | </table> | | </table> |
| | == Function == | | == Function == |
| - | [[http://www.uniprot.org/uniprot/4OT1_PSEPU 4OT1_PSEPU]] Catalyzes the ketonization of 2-hydroxymuconate stereoselectively to yield 2-oxo-3-hexenedioate.<ref>PMID:1339435</ref> | + | [https://www.uniprot.org/uniprot/4OT1_PSEPU 4OT1_PSEPU] Catalyzes the ketonization of 2-hydroxymuconate stereoselectively to yield 2-oxo-3-hexenedioate.<ref>PMID:1339435</ref> |
| | <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: 2-hydroxymuconate tautomerase]] | + | [[Category: Large Structures]] |
| - | [[Category: Poddar, H]] | + | [[Category: Pseudomonas putida]] |
| - | [[Category: Thunnissen, A M.W H]] | + | [[Category: Poddar H]] |
| - | [[Category: 4-oxalocrotonate tautomerase]] | + | [[Category: Thunnissen AMWH]] |
| - | [[Category: Beta-alpha-beta structural motif]]
| + | |
| - | [[Category: Isomerase]]
| + | |
| - | [[Category: Tautomerase superfamily]]
| + | |
| Structural highlights
Function
4OT1_PSEPU Catalyzes the ketonization of 2-hydroxymuconate stereoselectively to yield 2-oxo-3-hexenedioate.[1]
Publication Abstract from PubMed
The Michael-type addition reaction is widely used in organic synthesis for carbon-carbon bond formation. However, biocatalytic methodologies for this type of reaction are scarce, which is related to the fact that enzymes naturally catalysing carbon-carbon bond-forming Michael-type additions are rare. A promising template to develop new biocatalysts for carbon-carbon bond formation is the enzyme 4-oxalocrotonate tautomerase, which exhibits promiscuous Michael-type addition activity. Here we present mutability landscapes for the expression, tautomerase and Michael-type addition activities, and enantioselectivity of 4-oxalocrotonate tautomerase. These maps of neutral, beneficial and detrimental amino acids for each residue position and enzyme property provide detailed insight into sequence-function relationships. This offers exciting opportunities for enzyme engineering, which is illustrated by the redesign of 4-oxalocrotonate tautomerase into two enantiocomplementary 'Michaelases'. These 'Michaelases' catalyse the asymmetric addition of acetaldehyde to various nitroolefins, providing access to both enantiomers of gamma-nitroaldehydes, which are important precursors for pharmaceutically active gamma-aminobutyric acid derivatives.
Using mutability landscapes of a promiscuous tautomerase to guide the engineering of enantioselective Michaelases.,van der Meer JY, Poddar H, Baas BJ, Miao Y, Rahimi M, Kunzendorf A, van Merkerk R, Tepper PG, Geertsema EM, Thunnissen AM, Quax WJ, Poelarends GJ Nat Commun. 2016 Mar 8;7:10911. doi: 10.1038/ncomms10911. PMID:26952338[2]
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.
References
- ↑ Chen LH, Kenyon GL, Curtin F, Harayama S, Bembenek ME, Hajipour G, Whitman CP. 4-Oxalocrotonate tautomerase, an enzyme composed of 62 amino acid residues per monomer. J Biol Chem. 1992 Sep 5;267(25):17716-21. PMID:1339435
- ↑ van der Meer JY, Poddar H, Baas BJ, Miao Y, Rahimi M, Kunzendorf A, van Merkerk R, Tepper PG, Geertsema EM, Thunnissen AM, Quax WJ, Poelarends GJ. Using mutability landscapes of a promiscuous tautomerase to guide the engineering of enantioselective Michaelases. Nat Commun. 2016 Mar 8;7:10911. doi: 10.1038/ncomms10911. PMID:26952338 doi:http://dx.doi.org/10.1038/ncomms10911
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