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| | <StructureSection load='3uoy' size='340' side='right'caption='[[3uoy]], [[Resolution|resolution]] 2.00Å' scene=''> | | <StructureSection load='3uoy' size='340' side='right'caption='[[3uoy]], [[Resolution|resolution]] 2.00Å' scene=''> |
| | == Structural highlights == | | == Structural highlights == |
| - | <table><tr><td colspan='2'>[[3uoy]] is a 2 chain structure with sequence from [https://en.wikipedia.org/wiki/"bacillus_fluorescens_putidus"_flugge_1886 "bacillus fluorescens putidus" flugge 1886]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=3UOY OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=3UOY FirstGlance]. <br> | + | <table><tr><td colspan='2'>[[3uoy]] is a 2 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=3UOY OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=3UOY FirstGlance]. <br> |
| - | </td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=FAD:FLAVIN-ADENINE+DINUCLEOTIDE'>FAD</scene>, <scene name='pdbligand=NA:SODIUM+ION'>NA</scene>, <scene name='pdbligand=NAP:NADP+NICOTINAMIDE-ADENINE-DINUCLEOTIDE+PHOSPHATE'>NAP</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]] 2Å</td></tr> |
| - | <tr id='related'><td class="sblockLbl"><b>[[Related_structure|Related:]]</b></td><td class="sblockDat"><div style='overflow: auto; max-height: 3em;'>[[3uov|3uov]], [[3uox|3uox]], [[3uoz|3uoz]], [[3up4|3up4]], [[3up5|3up5]]</div></td></tr> | + | <tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=FAD:FLAVIN-ADENINE+DINUCLEOTIDE'>FAD</scene>, <scene name='pdbligand=NA:SODIUM+ION'>NA</scene>, <scene name='pdbligand=NAP:NADP+NICOTINAMIDE-ADENINE-DINUCLEOTIDE+PHOSPHATE'>NAP</scene></td></tr> |
| - | <tr id='gene'><td class="sblockLbl"><b>[[Gene|Gene:]]</b></td><td class="sblockDat">otemo ([https://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?mode=Info&srchmode=5&id=303 "Bacillus fluorescens putidus" Flugge 1886])</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=3uoy FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=3uoy OCA], [https://pdbe.org/3uoy PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=3uoy RCSB], [https://www.ebi.ac.uk/pdbsum/3uoy PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=3uoy 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=3uoy FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=3uoy OCA], [https://pdbe.org/3uoy PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=3uoy RCSB], [https://www.ebi.ac.uk/pdbsum/3uoy PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=3uoy ProSAT]</span></td></tr> |
| | </table> | | </table> |
| - | <div style="background-color:#fffaf0;">
| + | == Function == |
| - | == Publication Abstract from PubMed == | + | [https://www.uniprot.org/uniprot/OTEMO_PSEPU OTEMO_PSEPU] Involved in the degradation of (+)-camphor. Catalyzes the lactonization of 2-oxo-delta(3)-4,5, 5-trimethylcyclopentenylacetyl-CoA (OT-CoA), a key intermediate in the metabolism of camphor. 2-Oxocyclopentyl ethyl acetate is also a good substrate, as is 2-oxocyclohexyl ethyl acetate and methyl-substituted cyclohexanones, but free acid is a poor substrate.<ref>PMID:22267661</ref> <ref>PMID:22286514</ref> <ref>PMID:6848481</ref> |
| - | A dimeric Baeyer-Villiger monooxygenase (BVMO) catalyzing the lactonization of 2-oxo-Delta(3)-4,5,5-trimethylcyclopentenylacetyl-CoA, a key intermediate in the metabolism of camphor by Pseudomonas putida ATCC 17453 had been initially characterized in 1983 by Trudgill and co-workers (H.J. Ougham, D.G. Taylor, and P.W. Trudgill, J. Bacteriol. 153:140-152, 1983). Here we have cloned and overexpressed the 2-oxo-Delta(3)-4,5,5-trimethylcyclopentenylacetyl-CoA monooxygenase (OTEMO) in Escherichia coli, and determined its three-dimensional structure with bound FAD at 1.95 A resolution as well as with bound FAD and NADP(+) at 2.0 A resolution. OTEMO represents the first homodimeric type 1 BVMO structure bound to FAD/NADP(+). Comparison of several crystal forms of OTEMO bound to FAD and NADP(+) revealed conformational plasticity of several loop regions, some of which have been implicated as contributing to the substrate specificity profile of structurally-related BVMOs. Substrate specificity studies confirmed that the 2-oxo-Delta(3)-4,5,5-trimethylcyclopentenylacetic acid coenzyme A ester is preferred over the free acid. However, the catalytic efficiency (k(cat)/K(m)) favors 2-n-hexyl cyclopentanone (4.3 x 10(5) M(-1)s(-1)) as a substrate, although its affinity (K(m) = 32 muM) was lower than that of the CoA-activated substrate (18 muM). In whole cell biotransformation experiments, OTEMO showed a unique enantiocomplementarity to the action of the prototypical cyclohexanone monooxygenase (CHMO), and appeared to be particularly useful for the oxidation of 4-substituted cyclohexanones. Overall, this work expands our understanding of the molecular structure and mechanistic complexity of the type 1 family of BVMOs as well as expanding the catalytic repertoire of one of its original members.
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| - | Camphor Pathway 2-Oxo-{Delta}3-4,5,5-trimethylcyclopentenylacetyl-CoA Monooxygenase of Pseudomonas putida ATCC 17453: Cloning, Baeyer-Villiger Biooxidations, and Structures.,Leisch H, Shi R, Grosse S, Morley K, Bergeron H, Cygler M, Iwaki H, Hasegawa Y, Lau PC Appl Environ Microbiol. 2012 Jan 20. PMID:22267661<ref>PMID:22267661</ref>
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| - | | + | |
| - | From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br>
| + | |
| - | </div> | + | |
| - | <div class="pdbe-citations 3uoy" style="background-color:#fffaf0;"></div> | + | |
| | == References == | | == References == |
| | <references/> | | <references/> |
| | __TOC__ | | __TOC__ |
| | </StructureSection> | | </StructureSection> |
| - | [[Category: Bacillus fluorescens putidus flugge 1886]] | |
| | [[Category: Large Structures]] | | [[Category: Large Structures]] |
| - | [[Category: Cygler, M]] | + | [[Category: Pseudomonas putida]] |
| - | [[Category: Lau, P]] | + | [[Category: Cygler M]] |
| - | [[Category: Matte, A]] | + | [[Category: Lau P]] |
| - | [[Category: Shi, R]] | + | [[Category: Matte A]] |
| - | [[Category: Baeyer-villiger monooxygenase]]
| + | [[Category: Shi R]] |
| - | [[Category: Oxidoreductase]]
| + | |
| Structural highlights
Function
OTEMO_PSEPU Involved in the degradation of (+)-camphor. Catalyzes the lactonization of 2-oxo-delta(3)-4,5, 5-trimethylcyclopentenylacetyl-CoA (OT-CoA), a key intermediate in the metabolism of camphor. 2-Oxocyclopentyl ethyl acetate is also a good substrate, as is 2-oxocyclohexyl ethyl acetate and methyl-substituted cyclohexanones, but free acid is a poor substrate.[1] [2] [3]
References
- ↑ Leisch H, Shi R, Grosse S, Morley K, Bergeron H, Cygler M, Iwaki H, Hasegawa Y, Lau PC. Camphor Pathway 2-Oxo-{Delta}3-4,5,5-trimethylcyclopentenylacetyl-CoA Monooxygenase of Pseudomonas putida ATCC 17453: Cloning, Baeyer-Villiger Biooxidations, and Structures. Appl Environ Microbiol. 2012 Jan 20. PMID:22267661 doi:10.1128/AEM.07694-11
- ↑ Kadow M, Loschinski K, Sass S, Schmidt M, Bornscheuer UT. Completing the series of BVMOs involved in camphor metabolism of Pseudomonas putida NCIMB 10007 by identification of the two missing genes, their functional expression in E. coli, and biochemical characterization. Appl Microbiol Biotechnol. 2012 Oct;96(2):419-29. PMID:22286514 doi:http://dx.doi.org/10.1007/s00253-011-3859-1
- ↑ Ougham HJ, Taylor DG, Trudgill PW. Camphor revisited: involvement of a unique monooxygenase in metabolism of 2-oxo-delta 3-4,5,5-trimethylcyclopentenylacetic acid by Pseudomonas putida. J Bacteriol. 1983 Jan;153(1):140-52. PMID:6848481 doi:10.1128/jb.153.1.140-152.1983
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