2d0d
From Proteopedia
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|PDB= 2d0d |SIZE=350|CAPTION= <scene name='initialview01'>2d0d</scene>, resolution 1.65Å | |PDB= 2d0d |SIZE=350|CAPTION= <scene name='initialview01'>2d0d</scene>, resolution 1.65Å | ||
|SITE= | |SITE= | ||
| - | |LIGAND= <scene name='pdbligand=CL:CHLORIDE+ION'>CL</scene> | + | |LIGAND= <scene name='pdbligand=CL:CHLORIDE+ION'>CL</scene>, <scene name='pdbligand=PO4:PHOSPHATE+ION'>PO4</scene> |
| - | |ACTIVITY= [http://en.wikipedia.org/wiki/2-hydroxymuconate-semialdehyde_hydrolase 2-hydroxymuconate-semialdehyde hydrolase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=3.7.1.9 3.7.1.9] | + | |ACTIVITY= <span class='plainlinks'>[http://en.wikipedia.org/wiki/2-hydroxymuconate-semialdehyde_hydrolase 2-hydroxymuconate-semialdehyde hydrolase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=3.7.1.9 3.7.1.9] </span> |
|GENE= CUMD ([http://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?mode=Info&srchmode=5&id=294 Pseudomonas fluorescens]) | |GENE= CUMD ([http://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?mode=Info&srchmode=5&id=294 Pseudomonas fluorescens]) | ||
| + | |DOMAIN= | ||
| + | |RELATEDENTRY=[[1iup|1IUP]] | ||
| + | |RESOURCES=<span class='plainlinks'>[http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=2d0d FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=2d0d OCA], [http://www.ebi.ac.uk/pdbsum/2d0d PDBsum], [http://www.rcsb.org/pdb/explore.do?structureId=2d0d RCSB]</span> | ||
}} | }} | ||
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[[Category: Shoun, H.]] | [[Category: Shoun, H.]] | ||
[[Category: Wakagi, T.]] | [[Category: Wakagi, T.]] | ||
| - | [[Category: CL]] | ||
| - | [[Category: PO4]] | ||
[[Category: alpha/beta-hydrolase]] | [[Category: alpha/beta-hydrolase]] | ||
[[Category: beta-ketolase]] | [[Category: beta-ketolase]] | ||
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[[Category: substrate specificity]] | [[Category: substrate specificity]] | ||
| - | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on | + | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Mon Mar 31 02:28:38 2008'' |
Revision as of 23:28, 30 March 2008
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| , resolution 1.65Å | |||||||
|---|---|---|---|---|---|---|---|
| Ligands: | , | ||||||
| Gene: | CUMD (Pseudomonas fluorescens) | ||||||
| Activity: | 2-hydroxymuconate-semialdehyde hydrolase, with EC number 3.7.1.9 | ||||||
| Related: | 1IUP
| ||||||
| Resources: | FirstGlance, OCA, PDBsum, RCSB | ||||||
| Coordinates: | save as pdb, mmCIF, xml | ||||||
Crystal Structure of a Meta-cleavage Product Hydrolase (CumD) A129V Mutant
Overview
The meta-cleavage product hydrolase from Pseudomonas fluorescens IP01 (CumD) hydrolyzes 2-hydroxy-6-oxo-7-methylocta-2,4-dienoate (6-isopropyl HODA) in the cumene (isopropylbenzene) degradation pathway. To modulate the substrate specificity and catalytic efficiency of CumD toward substrates derived from monocyclic aromatic compounds, we constructed the CumD mutants, A129V, I199V, and V227I, as well as four types of double and triple mutants. Toward substrates with smaller side chains (e.g. 2-hydroxy-6-oxohepta-2,4-dienoate; 6-ethyl-HODA), the k(cat)/K(m) values of the single mutants were 4.2-11 fold higher than that of the wild type enzyme and 1.8-4.7 fold higher than that of the meta-cleavage product hydrolase from Pseudomonas putida F1 (TodF). The A129V mutant showed the highest k(cat)/K(m) value for 2-hydroxy-6-oxohepta-2,4-dienoate (6-ethyl-HODA). The crystal structure of the A129V mutant was determined at 1.65 A resolution, enabling location of the Ogamma atom of the Ser103 side chain. A chloride ion was bound to the oxyanion hole of the active site, and mutant enzymes at the residues forming this site were also examined. The k(cat) values of Ser34 mutants were decreased 2.9-65 fold, suggesting that the side chain of Ser34 supports catalysis by stabilizing the anionic oxygen of the proposed intermediate state (gem-diolate). This is the first crystal structure determination of CumD in an active form, with the Ser103 residue, one of the catalytically essential "triad", being intact.
About this Structure
2D0D is a Single protein structure of sequence from Pseudomonas fluorescens. Full crystallographic information is available from OCA.
Reference
Improving the catalytic efficiency of a meta-cleavage product hydrolase (CumD) from Pseudomonas fluorescens IP01., Jun SY, Fushinobu S, Nojiri H, Omori T, Shoun H, Wakagi T, Biochim Biophys Acta. 2006 Jul;1764(7):1159-66. Epub 2006 Jun 7. PMID:16844437
Page seeded by OCA on Mon Mar 31 02:28:38 2008
Categories: 2-hydroxymuconate-semialdehyde hydrolase | Pseudomonas fluorescens | Single protein | Fushinobu, S. | Jun, S Y. | Nojiri, H. | Omori, T. | Shoun, H. | Wakagi, T. | Alpha/beta-hydrolase | Beta-ketolase | Cumene degradation | Meta-cleavage product hydrolase | Pcb | Polychlorinated biphenyl degradation | Substrate specificity
