PDBsum entry 1phh

Oxidoreductase

PDB id: 1phh

Contents

Protein chain

394 a.a. *

Ligands

FAD

DHB

Waters ×273

* Residue conservation analysis

PDB id:

1phh

Name:

Oxidoreductase

Title:

Crystal structure of p-hydroxybenzoate hydroxylase complexed with its reaction product 3,4-dihydroxybenzoate

Structure:

P-hydroxybenzoate hydroxylase. Chain: a. Engineered: yes

Source:

Pseudomonas fluorescens. Organism_taxid: 294

Biol. unit:

Dimer (from PQS)

Resolution:

2.30Å R-factor: 0.193

Authors:

H.A.Schreuder,J.Drenth

Key ref:

H.A.Schreuder et al. (1988). Crystal structure of p-hydroxybenzoate hydroxylase complexed with its reaction product 3,4-dihydroxybenzoate. J Mol Biol, 199, 637-648. PubMed id: 3351945

Date:

04-Nov-87 Release date: 16-Apr-88

Protein chain

P00438  (PHHY_PSEFL) -  p-hydroxybenzoate hydroxylase from Pseudomonas fluorescens

Seq: 394 a.a.

Struc: 394 a.a.

Enzyme reactions

• Enzyme class: E.C.1.14.13.2 - 4-hydroxybenzoate 3-monooxygenase.
• Pathway: Benzoate Metabolism
• Reaction: 4-hydroxybenzoate + NADPH + O2 + H⁺ = 3,4-dihydroxybenzoate + NADP⁺ + H2O

4-hydroxybenzoate + NADPH + O2 + H(+) = 3,4-dihydroxybenzoate (Bound ligand (Het Group name = DHB) corresponds exactly) + NADP(+) + H2O

• Cofactor: FAD

FAD (Bound ligand (Het Group name = FAD) corresponds exactly)

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

reference

J Mol Biol 199:637-648 (1988)

PubMed id: 3351945

Crystal structure of p-hydroxybenzoate hydroxylase complexed with its reaction product 3,4-dihydroxybenzoate.

H.A.Schreuder, J.M.van der Laan, W.G.Hol, J.Drenth.

ABSTRACT

Crystals of the flavin-containing enzyme p-hydroxybenzoate hydroxylase (PHBHase) complexed with its reaction product were investigated in order to obtain insight into the catalytic cycle of this enzyme involving two substrates and two cofactors. PHBHase was crystallized initially with its substrate, p-hydroxybenzoate and the substrate was then converted into the product 3,4-dihydroxybenzoate by allowing the catalytic reaction to proceed in the crystals. In addition, crystals were soaked in mother liquor containing a high concentration of this product. Data up to 2.3 A (1 A = 0.1 nm) were collected by the oscillation method and the structure of the enzyme product complex was refined by alternate restrained least-squares procedures and model building by computer graphics techniques. A total of 273 solvent molecules could be located, four of them being presumably sulfate ions. The R-factor for 14,339 reflections between 6.0 A and 2.3 A is 19.3%. The 3-hydroxyl group of the product introduced by the enzyme is clearly visible in the electron density, showing unambiguously which carbon atom of the substrate is hydroxylated. A clear picture of the hydroxylation site is obtained. The plane of the product is rotated 21 degrees with respect to the plane of the substrate in the current model of enzyme-substrate complex. The 4-hydroxyl group of the product is hydrogen bonded to the hydroxyl group of Tyr201, its carboxyl group is interacting with the side-chains of Tyr222, Arg214 and Ser212, while the newly introduced 3-hydroxyl group makes a hydrogen bond with the backbone carbonyl oxygen of Pro293.