|
|
| (One intermediate revision not shown.) |
| Line 1: |
Line 1: |
| | | | |
| | ==Crystal structure of the Thermotoga maritima acetyl esterase (TM0077) complex with a substrate analog== | | ==Crystal structure of the Thermotoga maritima acetyl esterase (TM0077) complex with a substrate analog== |
| - | <StructureSection load='5jib' size='340' side='right' caption='[[5jib]], [[Resolution|resolution]] 1.86Å' scene=''> | + | <StructureSection load='5jib' size='340' side='right'caption='[[5jib]], [[Resolution|resolution]] 1.86Å' scene=''> |
| | == Structural highlights == | | == Structural highlights == |
| - | <table><tr><td colspan='2'>[[5jib]] is a 6 chain structure. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=5JIB OCA]. For a <b>guided tour on the structure components</b> use [http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=5JIB FirstGlance]. <br> | + | <table><tr><td colspan='2'>[[5jib]] is a 6 chain structure with sequence from [https://en.wikipedia.org/wiki/Thermotoga_maritima_MSB8 Thermotoga maritima MSB8]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=5JIB OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=5JIB FirstGlance]. <br> |
| - | </td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat"><scene name='pdbligand=OIA:[(3S)-2-OXO-2,3-DIHYDRO-1H-INDOL-3-YL]ACETIC+ACID'>OIA</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]] 1.86Å</td></tr> |
| - | <tr id='related'><td class="sblockLbl"><b>[[Related_structure|Related:]]</b></td><td class="sblockDat">[[5fdf|5fdf]], [[5hfn|5hfn]], [[3m82|3m82]], [[3m83|3m83]]</td></tr> | + | <tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=OIA:[(3S)-2-OXO-2,3-DIHYDRO-1H-INDOL-3-YL]ACETIC+ACID'>OIA</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=5jib FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=5jib OCA], [http://pdbe.org/5jib PDBe], [http://www.rcsb.org/pdb/explore.do?structureId=5jib RCSB], [http://www.ebi.ac.uk/pdbsum/5jib PDBsum], [http://prosat.h-its.org/prosat/prosatexe?pdbcode=5jib ProSAT]</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=5jib FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=5jib OCA], [https://pdbe.org/5jib PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=5jib RCSB], [https://www.ebi.ac.uk/pdbsum/5jib PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=5jib ProSAT]</span></td></tr> |
| | </table> | | </table> |
| | == Function == | | == Function == |
| - | [[http://www.uniprot.org/uniprot/CAH_THEMA CAH_THEMA]] Esterase that removes acetyl groups from a number of O-acetylated small substrates, such as acetylated xylose, short xylo-oligosaccharides and cephalosporin C. Has no activity towards polymeric acetylated xylan, 4-methylumbelliferyl acetate or alpha-naphthyl acetate. Able to catalyze rapid hydrolysis of a range of substrates preferably with acetate groups, independent of the alcohol moiety. Exhibits a narrow selectivity for short chain acyl esters (C2-C3). Displays broad substrate specificity by hydrolyzing acetate at 2, 3, and 4 positions of 4-nitrophenyl-beta-D-xylopyranoside (pNP-Xyl) with similar efficiency. Cannot cleave amide linkages.<ref>PMID:21255309</ref> <ref>PMID:22411095</ref> <ref>PMID:22659119</ref> | + | [https://www.uniprot.org/uniprot/CAH_THEMA CAH_THEMA] Esterase that removes acetyl groups from a number of O-acetylated small substrates, such as acetylated xylose, short xylo-oligosaccharides and cephalosporin C. Has no activity towards polymeric acetylated xylan, 4-methylumbelliferyl acetate or alpha-naphthyl acetate. Able to catalyze rapid hydrolysis of a range of substrates preferably with acetate groups, independent of the alcohol moiety. Exhibits a narrow selectivity for short chain acyl esters (C2-C3). Displays broad substrate specificity by hydrolyzing acetate at 2, 3, and 4 positions of 4-nitrophenyl-beta-D-xylopyranoside (pNP-Xyl) with similar efficiency. Cannot cleave amide linkages.<ref>PMID:21255309</ref> <ref>PMID:22411095</ref> <ref>PMID:22659119</ref> |
| | <div style="background-color:#fffaf0;"> | | <div style="background-color:#fffaf0;"> |
| | == Publication Abstract from PubMed == | | == Publication Abstract from PubMed == |
| Line 23: |
Line 23: |
| | __TOC__ | | __TOC__ |
| | </StructureSection> | | </StructureSection> |
| - | [[Category: Manoj, N]] | + | [[Category: Large Structures]] |
| - | [[Category: Carbohydrate metabolism]] | + | [[Category: Thermotoga maritima MSB8]] |
| - | [[Category: Cephalosporin deacetylase]] | + | [[Category: Manoj N]] |
| - | [[Category: Hydrolase]]
| + | |
| - | [[Category: Rossmann fold]]
| + | |
| Structural highlights
Function
CAH_THEMA Esterase that removes acetyl groups from a number of O-acetylated small substrates, such as acetylated xylose, short xylo-oligosaccharides and cephalosporin C. Has no activity towards polymeric acetylated xylan, 4-methylumbelliferyl acetate or alpha-naphthyl acetate. Able to catalyze rapid hydrolysis of a range of substrates preferably with acetate groups, independent of the alcohol moiety. Exhibits a narrow selectivity for short chain acyl esters (C2-C3). Displays broad substrate specificity by hydrolyzing acetate at 2, 3, and 4 positions of 4-nitrophenyl-beta-D-xylopyranoside (pNP-Xyl) with similar efficiency. Cannot cleave amide linkages.[1] [2] [3]
Publication Abstract from PubMed
The carbohydrate esterase family 7 (CE7) members are acetyl esterases that possess unusual substrate specificity for cephalosporin C and 7-amino-cephalosporanic acid. This family containing the alpha/beta hydrolase fold has a distinctive substrate profile that allows it to carry out hydrolysis of esters containing diverse alcohol moieties while maintaining narrow specificity for an acetate ester. Here we investigate the structural basis of this preference for small acyl groups using the crystal structure of the thermostable Thermotoga maritima CE7 acetyl esterase (TmAcE) complexed with a non-cognate substrate analog. The structure determined at 1.86 A resolution provides direct evidence for the location of the largely hydrophobic and rigid substrate binding pocket in this family. Furthermore, a three-helix insertion domain near the catalytic machinery shapes the substrate binding site. The structure reveals two residues (Pro228 and Ile276) which constitute a hydrophobic rigid binding surface for the acyl group of the ester and thus restricts the size of the acyl group that be accommodated. In combination with previous literature on kinetic properties of the enzyme, our studies suggest that these residues determine the unique specificity of the TmAcE for short straight chain esters. The structure provides a template for focused attempts to engineer the CE7 enzymes for enhanced stability, selectivity or activity for biocatalytic applications.
Crystal structure of Thermotoga maritima acetyl esterase complex with a substrate analog: Insights into the distinctive substrate specificity in the CE7 carbohydrate esterase family.,Singh MK, Manoj N Biochem Biophys Res Commun. 2016 Jul 22;476(2):63-8. doi:, 10.1016/j.bbrc.2016.05.061. Epub 2016 May 12. PMID:27181355[4]
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.
References
- ↑ Drzewiecki K, Angelov A, Ballschmiter M, Tiefenbach KJ, Sterner R, Liebl W. Hyperthermostable acetyl xylan esterase. Microb Biotechnol. 2010 Jan;3(1):84-92. doi: 10.1111/j.1751-7915.2009.00150.x., Epub 2009 Sep 18. PMID:21255309 doi:http://dx.doi.org/10.1111/j.1751-7915.2009.00150.x
- ↑ Levisson M, Han GW, Deller MC, Xu Q, Biely P, Hendriks S, Ten Eyck LF, Flensburg C, Roversi P, Miller MD, McMullan D, von Delft F, Kreusch A, Deacon AM, van der Oost J, Lesley SA, Elsliger MA, Kengen SW, Wilson IA. Functional and structural characterization of a thermostable acetyl esterase from Thermotoga maritima. Proteins. 2012 Jan 27. doi: 10.1002/prot.24041. PMID:22411095 doi:10.1002/prot.24041
- ↑ Hedge MK, Gehring AM, Adkins CT, Weston LA, Lavis LD, Johnson RJ. The structural basis for the narrow substrate specificity of an acetyl esterase from Thermotoga maritima. Biochim Biophys Acta. 2012 Sep;1824(9):1024-30. doi:, 10.1016/j.bbapap.2012.05.009. Epub 2012 Jun 1. PMID:22659119 doi:http://dx.doi.org/10.1016/j.bbapap.2012.05.009
- ↑ Singh MK, Manoj N. Crystal structure of Thermotoga maritima acetyl esterase complex with a substrate analog: Insights into the distinctive substrate specificity in the CE7 carbohydrate esterase family. Biochem Biophys Res Commun. 2016 Jul 22;476(2):63-8. doi:, 10.1016/j.bbrc.2016.05.061. Epub 2016 May 12. PMID:27181355 doi:http://dx.doi.org/10.1016/j.bbrc.2016.05.061
|
