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| | <StructureSection load='6iyl' size='340' side='right'caption='[[6iyl]], [[Resolution|resolution]] 2.56Å' scene=''> | | <StructureSection load='6iyl' size='340' side='right'caption='[[6iyl]], [[Resolution|resolution]] 2.56Å' scene=''> |
| | == Structural highlights == | | == Structural highlights == |
| - | <table><tr><td colspan='2'>[[6iyl]] is a 2 chain structure with sequence from [http://en.wikipedia.org/wiki/Escherichia_coli_1303 Escherichia coli 1303]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=6IYL OCA]. For a <b>guided tour on the structure components</b> use [http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=6IYL FirstGlance]. <br> | + | <table><tr><td colspan='2'>[[6iyl]] is a 2 chain structure with sequence from [https://en.wikipedia.org/wiki/Escherichia_coli_1303 Escherichia coli 1303]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=6IYL OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=6IYL FirstGlance]. <br> |
| - | </td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat"><scene name='pdbligand=B1X:5-O-[(3-cyanobenzene-1-carbonyl)sulfamoyl]adenosine'>B1X</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]] 2.56Å</td></tr> |
| - | <tr id='gene'><td class="sblockLbl"><b>[[Gene|Gene:]]</b></td><td class="sblockDat">entE, EC1303_c05680 ([http://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?mode=Info&srchmode=5&id=745156 Escherichia coli 1303])</td></tr> | + | <tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=B1X:5-O-[(3-cyanobenzene-1-carbonyl)sulfamoyl]adenosine'>B1X</scene></td></tr> |
| - | <tr id='activity'><td class="sblockLbl"><b>Activity:</b></td><td class="sblockDat"><span class='plainlinks'>[http://en.wikipedia.org/wiki/(2,3-dihydroxybenzoyl)adenylate_synthase (2,3-dihydroxybenzoyl)adenylate synthase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=2.7.7.58 2.7.7.58] </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=6iyl FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=6iyl OCA], [https://pdbe.org/6iyl PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=6iyl RCSB], [https://www.ebi.ac.uk/pdbsum/6iyl PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=6iyl ProSAT]</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=6iyl FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=6iyl OCA], [http://pdbe.org/6iyl PDBe], [http://www.rcsb.org/pdb/explore.do?structureId=6iyl RCSB], [http://www.ebi.ac.uk/pdbsum/6iyl PDBsum], [http://prosat.h-its.org/prosat/prosatexe?pdbcode=6iyl ProSAT]</span></td></tr> | + | |
| | </table> | | </table> |
| | + | == Function == |
| | + | [https://www.uniprot.org/uniprot/ENTE_ECOLI ENTE_ECOLI] Activates the carboxylate group of 2,3-dihydroxy-benzoate (2,3-DHB), via ATP-dependent PPi exchange reactions, to the acyladenylate. Then, catalyzes the acylation of holo-EntB with 2,3-DHB adenylate, preparing that molecule for amide bond formation with L-serine. |
| | <div style="background-color:#fffaf0;"> | | <div style="background-color:#fffaf0;"> |
| | == Publication Abstract from PubMed == | | == Publication Abstract from PubMed == |
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| | [[Category: Escherichia coli 1303]] | | [[Category: Escherichia coli 1303]] |
| | [[Category: Large Structures]] | | [[Category: Large Structures]] |
| - | [[Category: Ishikawa, F]] | + | [[Category: Ishikawa F]] |
| - | [[Category: Miyanaga, A]] | + | [[Category: Miyanaga A]] |
| - | [[Category: Adenylation]]
| + | |
| - | [[Category: Ligase]]
| + | |
| - | [[Category: Non-ribosomal peptide biosynthesis]]
| + | |
| Structural highlights
Function
ENTE_ECOLI Activates the carboxylate group of 2,3-dihydroxy-benzoate (2,3-DHB), via ATP-dependent PPi exchange reactions, to the acyladenylate. Then, catalyzes the acylation of holo-EntB with 2,3-DHB adenylate, preparing that molecule for amide bond formation with L-serine.
Publication Abstract from PubMed
Adenylation (A) domains act as the gatekeepers of non-ribosomal peptide synthetases (NRPSs) ensuring the activation and thioesterification of the correct amino acid/aryl acid building blocks. Aryl acid building blocks are most commonly observed in iron-chelating siderophores, but are not limited to them. The non-ribosomal codes toward aryl acid substrates are poorly understood. Very little is known about the reprogramming of aryl acid A-domains. Here we show that a single asparagine-to-glycine mutation in an aryl acid A-domain creates novel enzyme specificities toward a wide range of non-native aryl acids. The engineered catalyst is capable of activating the non-native aryl acids functionalized with nitro, cyano, bromo, and iodo, even though no enzymatic activity of wild-type enzyme was observed toward these substrates. Co-crystal structures with non-hydrolysable aryl-AMP analogues revealed the origins of substrate promiscuity expansion, highlighting an enlarged substrate binding pocket of the enzyme. Our finding may be exploited to produce diversified aryl acid-containing natural products and serve as a template for further directed evolution in combinatorial biosynthesis.
An engineered aryl acid adenylation domain with a capacious active site microenvironment.,Ishikawa F, Miyanaga A, Kitayama H, Nakamura S, Nakanishi I, Kudo F, Eguchi T, Tanabe G Angew Chem Int Ed Engl. 2019 Apr 3. doi: 10.1002/anie.201900318. PMID:30945421[1]
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.
References
- ↑ Ishikawa F, Miyanaga A, Kitayama H, Nakamura S, Nakanishi I, Kudo F, Eguchi T, Tanabe G. An engineered aryl acid adenylation domain with a capacious active site microenvironment. Angew Chem Int Ed Engl. 2019 Apr 3. doi: 10.1002/anie.201900318. PMID:30945421 doi:http://dx.doi.org/10.1002/anie.201900318
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