3up5: Difference between revisions
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==Crystal Structure of OTEMO complex with FAD and NADP (form 4)== | |||
=== | <StructureSection load='3up5' size='340' side='right' caption='[[3up5]], [[Resolution|resolution]] 2.45Å' scene=''> | ||
== Structural highlights == | |||
<table><tr><td colspan='2'>[[3up5]] is a 2 chain structure with sequence from [http://en.wikipedia.org/wiki/Pseudomonas_putida Pseudomonas putida]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=3UP5 OCA]. For a <b>guided tour on the structure components</b> use [http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=3UP5 FirstGlance]. <br> | |||
</td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat"><scene name='pdbligand=FAD:FLAVIN-ADENINE+DINUCLEOTIDE'>FAD</scene>, <scene name='pdbligand=NAP:NADP+NICOTINAMIDE-ADENINE-DINUCLEOTIDE+PHOSPHATE'>NAP</scene></td></tr> | |||
<tr id='related'><td class="sblockLbl"><b>[[Related_structure|Related:]]</b></td><td class="sblockDat">[[3uov|3uov]], [[3uox|3uox]], [[3uoy|3uoy]], [[3uoz|3uoz]], [[3up4|3up4]]</td></tr> | |||
<tr id='gene'><td class="sblockLbl"><b>[[Gene|Gene:]]</b></td><td class="sblockDat">otemo ([http://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?mode=Info&srchmode=5&id=303 Pseudomonas putida])</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=3up5 FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=3up5 OCA], [http://www.rcsb.org/pdb/explore.do?structureId=3up5 RCSB], [http://www.ebi.ac.uk/pdbsum/3up5 PDBsum]</span></td></tr> | |||
</table> | |||
<div style="background-color:#fffaf0;"> | |||
== Publication Abstract from PubMed == | |||
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. | |||
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> | |||
== | From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br> | ||
</div> | |||
== References == | |||
<references/> | |||
__TOC__ | |||
</StructureSection> | |||
[[Category: Pseudomonas putida]] | [[Category: Pseudomonas putida]] | ||
[[Category: Cygler, M | [[Category: Cygler, M]] | ||
[[Category: Lau, P | [[Category: Lau, P]] | ||
[[Category: Matte, A | [[Category: Matte, A]] | ||
[[Category: Shi, R | [[Category: Shi, R]] | ||
[[Category: Baeyer-villiger monooxygenase]] | [[Category: Baeyer-villiger monooxygenase]] | ||
[[Category: Oxidoreductase]] | [[Category: Oxidoreductase]] | ||
