1IUN

meta-Cleavage product hydrolase from Pseudomonas fluorescens IP01 (CumD) S103A mutant hexagonal

Summary for 1IUN

• Entry DOI: 10.2210/pdb1iun/pdb
• Related: 1IUO 1IUP
• Descriptor: meta-Cleavage product hydrolase, ACETATE ION (3 entities in total)
• Functional Keywords: aromatic compounds, cumene, isopropylbenzene, meta-cleavage compound hydrolase, polychlorinated biphenyl degradation, pseudomonas fluorescens ip01, alpha/beta-hydrolase, substrate specificity, cumene degradation, pcb, beta-ketolase, hydrolase
• Biological source: Pseudomonas fluorescens
• Total number of polymer chains: 2
• Total formula weight: 63072.26

Authors

Fushinobu, S.,Saku, T.,Hidaka, M.,Jun, S.-Y.,Nojiri, H.,Yamane, H.,Shoun, H.,Omori, T.,Wakagi, T. (deposition date: 2002-03-06, release date: 2002-09-18, Last modification date: 2023-10-25)

Primary citation

Fushinobu, S.,Saku, T.,Hidaka, M.,Jun, S.-Y.,Nojiri, H.,Yamane, H.,Shoun, H.,Omori, T.,Wakagi, T.
Crystal structures of a meta-cleavage product hydrolase from Pseudomonas fluorescens IP01 (CumD) complexed with cleavage products
PROTEIN SCI., 11:2184-2195, 2002

PubMed Abstract: 2-Hydroxy-6-oxo-7-methylocta-2,4-dienoate hydrolase (CumD) from Pseudomonas fluorescens IP01 hydrolyzes a meta-cleavage product generated in the cumene (isopropylbenzene) degradation pathway. The crystal structures of the inactive S103A mutant of the CumD enzyme complexed with isobutyrate and acetate ions were determined at 1.6 and 2.0 A resolution, respectively. The isobutyrate and acetate ions were located at the same position in the active site, and occupied the site for a part of the hydrolysis product with CumD, which has the key determinant group for the substrate specificity of related hydrolases. One of the oxygen atoms of the carboxyl group of the isobutyrate ion was hydrogen bonded with a water molecule and His252. Another oxygen atom of the carboxyl group was situated in an oxyanion hole formed by the two main-chain N atoms. The isopropyl group of the isobutyric acid was recognized by the side-chains of the hydrophobic residues. The substrate-binding pocket of CumD was long, and the inhibition constants of various organic acids corresponded well to it. In comparison with the structure of BphD from Rhodococcus sp. RHA1, the structural basis for the substrate specificity of related hydrolases, is revealed.

PubMed: 12192074
DOI: 10.1110/ps.0209602

Experimental method

X-RAY DIFFRACTION (2.8 Å)