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| | ==Crystal structure of carbapenem synthase in complex with (3S,5S)-carbapenam== | | ==Crystal structure of carbapenem synthase in complex with (3S,5S)-carbapenam== |
| - | <StructureSection load='4oj8' size='340' side='right' caption='[[4oj8]], [[Resolution|resolution]] 2.10Å' scene=''> | + | <StructureSection load='4oj8' size='340' side='right'caption='[[4oj8]], [[Resolution|resolution]] 2.10Å' scene=''> |
| | == Structural highlights == | | == Structural highlights == |
| - | <table><tr><td colspan='2'>[[4oj8]] is a 3 chain structure with sequence from [http://en.wikipedia.org/wiki/Erwinia_carotovora_carotovora Erwinia carotovora carotovora]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=4OJ8 OCA]. For a <b>guided tour on the structure components</b> use [http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=4OJ8 FirstGlance]. <br> | + | <table><tr><td colspan='2'>[[4oj8]] is a 3 chain structure with sequence from [https://en.wikipedia.org/wiki/Pectobacterium_carotovorum_subsp._carotovorum Pectobacterium carotovorum subsp. carotovorum]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=4OJ8 OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=4OJ8 FirstGlance]. <br> |
| - | </td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat"><scene name='pdbligand=2TQ:(2S,5S)-7-OXO-1-AZABICYCLO[3.2.0]HEPTANE-2-CARBOXYLIC+ACID'>2TQ</scene>, <scene name='pdbligand=AKG:2-OXOGLUTARIC+ACID'>AKG</scene>, <scene name='pdbligand=FE2:FE+(II)+ION'>FE2</scene>, <scene name='pdbligand=GOL:GLYCEROL'>GOL</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]] 2.1Å</td></tr> |
| - | <tr id='gene'><td class="sblockLbl"><b>[[Gene|Gene:]]</b></td><td class="sblockDat">carC ([http://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?mode=Info&srchmode=5&id=555 Erwinia carotovora carotovora])</td></tr>
| + | <tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=2TQ:(2S,5S)-7-OXO-1-AZABICYCLO[3.2.0]HEPTANE-2-CARBOXYLIC+ACID'>2TQ</scene>, <scene name='pdbligand=AKG:2-OXOGLUTARIC+ACID'>AKG</scene>, <scene name='pdbligand=FE2:FE+(II)+ION'>FE2</scene>, <scene name='pdbligand=GOL:GLYCEROL'>GOL</scene></td></tr> |
| - | <tr id='activity'><td class="sblockLbl"><b>Activity:</b></td><td class="sblockDat"><span class='plainlinks'>[http://en.wikipedia.org/wiki/(5R)-carbapenem-3-carboxylate_synthase (5R)-carbapenem-3-carboxylate synthase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=1.14.20.3 1.14.20.3] </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=4oj8 FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=4oj8 OCA], [https://pdbe.org/4oj8 PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=4oj8 RCSB], [https://www.ebi.ac.uk/pdbsum/4oj8 PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=4oj8 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=4oj8 FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=4oj8 OCA], [http://www.rcsb.org/pdb/explore.do?structureId=4oj8 RCSB], [http://www.ebi.ac.uk/pdbsum/4oj8 PDBsum]</span></td></tr> | + | |
| | </table> | | </table> |
| | == Function == | | == Function == |
| - | [[http://www.uniprot.org/uniprot/CARC_PECCC CARC_PECCC]] Catalyzes the Fe(2+) and alpha-ketoglutarate-dependent conversion of (3S,5S)-carbapenam to (5R)-carbapenem, an essential step in carbapenem antibiotic biosynthesis.<ref>PMID:9402024</ref> <ref>PMID:12848554</ref> <ref>PMID:15174175</ref> <ref>PMID:12611886</ref> | + | [https://www.uniprot.org/uniprot/CARC_PECCC CARC_PECCC] Catalyzes the Fe(2+) and alpha-ketoglutarate-dependent conversion of (3S,5S)-carbapenam to (5R)-carbapenem, an essential step in carbapenem antibiotic biosynthesis.<ref>PMID:9402024</ref> <ref>PMID:12848554</ref> <ref>PMID:15174175</ref> <ref>PMID:12611886</ref> |
| | <div style="background-color:#fffaf0;"> | | <div style="background-color:#fffaf0;"> |
| | == Publication Abstract from PubMed == | | == Publication Abstract from PubMed == |
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| | From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br> | | From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br> |
| | </div> | | </div> |
| | + | <div class="pdbe-citations 4oj8" style="background-color:#fffaf0;"></div> |
| | == References == | | == References == |
| | <references/> | | <references/> |
| | __TOC__ | | __TOC__ |
| | </StructureSection> | | </StructureSection> |
| - | [[Category: Erwinia carotovora carotovora]] | + | [[Category: Large Structures]] |
| - | [[Category: Boal, A K]] | + | [[Category: Pectobacterium carotovorum subsp. carotovorum]] |
| - | [[Category: Rosenzweig, A C]] | + | [[Category: Boal AK]] |
| - | [[Category: 2-oxoglutaric acid]] | + | [[Category: Rosenzweig AC]] |
| - | [[Category: Antibiotic biosynthesis]]
| + | |
| - | [[Category: Carbapenam]]
| + | |
| - | [[Category: Iron]]
| + | |
| - | [[Category: Oxidoreductase]]
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| - | [[Category: Oxygenase]]
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| Structural highlights
Function
CARC_PECCC Catalyzes the Fe(2+) and alpha-ketoglutarate-dependent conversion of (3S,5S)-carbapenam to (5R)-carbapenem, an essential step in carbapenem antibiotic biosynthesis.[1] [2] [3] [4]
Publication Abstract from PubMed
The bicyclic beta-lactam/2-pyrrolidine precursor to all carbapenem antibiotics is biosynthesized by attachment of a carboxymethylene unit to C5 of L-proline followed by beta-lactam ring closure. Carbapenem synthase (CarC), an Fe(II) and 2-(oxo)glutarate (Fe/2OG)-dependent oxygenase, then inverts the C5 configuration. Here we report the structure of CarC in complex with its substrate and biophysical dissection of its reaction to reveal the stereoinversion mechanism. An Fe(IV)-oxo intermediate abstracts the hydrogen (H*) from C5, and tyrosine 165, a residue not visualized in the published structures of CarC lacking bound substrate, donates H* to the opposite face of the resultant radical. The reaction oxidizes the Fe(II) cofactor to Fe(III), limiting wild-type CarC to one turnover, but substitution of the H*-donating tyrosine disables stereoinversion and confers to CarC the capacity for catalytic substrate oxidation.
Mechanism of the C5 stereoinversion reaction in the biosynthesis of carbapenem antibiotics.,Chang WC, Guo Y, Wang C, Butch SE, Rosenzweig AC, Boal AK, Krebs C, Bollinger JM Jr Science. 2014 Mar 7;343(6175):1140-4. doi: 10.1126/science.1248000. PMID:24604200[5]
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.
References
- ↑ McGowan SJ, Sebaihia M, O'Leary S, Hardie KR, Williams P, Stewart GS, Bycroft BW, Salmond GP. Analysis of the carbapenem gene cluster of Erwinia carotovora: definition of the antibiotic biosynthetic genes and evidence for a novel beta-lactam resistance mechanism. Mol Microbiol. 1997 Nov;26(3):545-56. PMID:9402024
- ↑ Stapon A, Li R, Townsend CA. Carbapenem biosynthesis: confirmation of stereochemical assignments and the role of CarC in the ring stereoinversion process from L-proline. J Am Chem Soc. 2003 Jul 16;125(28):8486-93. PMID:12848554 doi:http://dx.doi.org/10.1021/ja034248a
- ↑ Sleeman MC, Smith P, Kellam B, Chhabra SR, Bycroft BW, Schofield CJ. Biosynthesis of carbapenem antibiotics: new carbapenam substrates for carbapenem synthase (CarC). Chembiochem. 2004 Jun 7;5(6):879-82. PMID:15174175 doi:http://dx.doi.org/10.1002/cbic.200300908
- ↑ Clifton IJ, Doan LX, Sleeman MC, Topf M, Suzuki H, Wilmouth RC, Schofield CJ. Crystal structure of carbapenem synthase (CarC). J Biol Chem. 2003 Jun 6;278(23):20843-50. Epub 2003 Feb 28. PMID:12611886 doi:10.1074/jbc.M213054200
- ↑ Chang WC, Guo Y, Wang C, Butch SE, Rosenzweig AC, Boal AK, Krebs C, Bollinger JM Jr. Mechanism of the C5 stereoinversion reaction in the biosynthesis of carbapenem antibiotics. Science. 2014 Mar 7;343(6175):1140-4. doi: 10.1126/science.1248000. PMID:24604200 doi:http://dx.doi.org/10.1126/science.1248000
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