PDBsum entry 2hml

Oxidoreductase

PDB id: 2hml

Contents

Protein chains

446 a.a. *

192 a.a. *

Ligands

SO4 ×5

FES

PEY

EDO ×17

Metals

_FE

Waters ×550

* Residue conservation analysis

PDB id:

2hml

Name:

Oxidoreductase

Title:

Crystal structure of the naphthalene 1,2-dioxygenase f352v mutant bound to phenanthrene.

Structure:

Naphthalene 1,2-dioxygenase alpha subunit. Chain: a. Synonym: naphthalene 1,2-dioxygenase isp alpha. Engineered: yes. Mutation: yes. Naphthalene 1,2-dioxygenase beta subunit. Chain: b. Synonym: naphthalene 1,2-dioxygenase isp beta. Engineered: yes

Source:

Pseudomonas sp.. Organism_taxid: 306. Gene: doxb. Expressed in: escherichia coli. Expression_system_taxid: 562. Gene: doxd.

Biol. unit:

Hexamer (from PDB file)

Resolution:

1.80Å R-factor: 0.165 R-free: 0.190

Authors:

D.J.Ferraro,A.L.Okerlund,J.C.Mowers,S.Ramaswamy

Key ref:

D.J.Ferraro et al. (2006). Structural basis for regioselectivity and stereoselectivity of product formation by naphthalene 1,2-dioxygenase. J Bacteriol, 188, 6986-6994. PubMed id: 16980501

Date:

11-Jul-06 Release date: 10-Oct-06

Protein chain

P0A111  (NDOB_PSEU8) -  Naphthalene 1,2-dioxygenase system, large oxygenase component from Pseudomonas sp. (strain C18)

Seq: 449 a.a.

Struc: 446 a.a.*

Protein chain

P0A113  (NDOC_PSEU8) -  Naphthalene 1,2-dioxygenase system, small oxygenase component from Pseudomonas sp. (strain C18)

Seq: 194 a.a.

Struc: 192 a.a.

* PDB and UniProt seqs differ at 1 residue position (black cross)

Enzyme reactions

• Enzyme class: Chain A: E.C.1.14.12.12 - naphthalene 1,2-dioxygenase.
• Pathway: Aromatic 1,2-Dioxygenases
• Reaction: naphthalene + NADH + O2 + H⁺ = (1R,2S)-1,2-dihydronaphthalene-1,2-diol + NAD⁺

naphthalene + NADH + O2 (Bound ligand (Het Group name = PEY) matches with 71.43% similarity) + H(+) = (1R,2S)-1,2-dihydronaphthalene-1,2-diol + NAD(+)

• Cofactor: Fe cation

Molecule diagrams generated from .mol files obtained from the KEGG ftp site

reference

J Bacteriol 188:6986-6994 (2006)

PubMed id: 16980501

Structural basis for regioselectivity and stereoselectivity of product formation by naphthalene 1,2-dioxygenase.

D.J.Ferraro, A.L.Okerlund, J.C.Mowers, S.Ramaswamy.

ABSTRACT

Rieske oxygenase (RO) systems are two- and three-component enzyme systems that catalyze the formation of cis-dihydrodiols from aromatic substrates. Degradation of pollutants in contaminated soil and generation of chiral synthons have been the major foci of RO research. Substrate specificity and product regio- and stereoselectivity have been shown to vary between individual ROs. While directed evolution methods for altering RO function have been successful in the past, rational engineering of these enzymes still poses a challenge due to the lack of structural understanding. Here we examine the structural changes induced by mutation of Phe-352 in naphthalene 1,2-dioxygenase from Pseudomonas sp. strain NCIB 9816-4 (NDO-O(9816-4)). Structures of the Phe-352-Val mutant in native form and in complex with phenanthrene and anthracene, along with those of wild-type NDO-O(9816-4) in complex with phenanthrene, anthracene, and 3-nitrotoluene, are presented. Phenanthrene was shown to bind in a different orientation in the Phe-352-Val mutant active site from that in the wild type, while anthracene was found to bind in similar positions in both enzymes. Two orientations of 3-nitrotoluene were observed, i.e., a productive and a nonproductive orientation. These orientations help explain why NDO-O(9816-4) forms different products from 3-nitrotoluene than those made from nitrobenzene dioxygenase. Comparison of these structures among themselves and with other known ROs bound to substrates reveals that the orientation of substrate binding at the active site is the primary determinant of product regio- and stereoselectivity.