PDBsum entry 3olu

Oxidoreductase

PDB id: 3olu

Contents

Protein chains

551 a.a.

Ligands

NAG-NAG ×2

NAG-NAG-MAN ×2

EDO ×14

1AG ×2

COH ×2

NAG ×2

BOG

Waters ×666

PDB id:

3olu

Name:

Oxidoreductase

Title:

X-ray crystal structure of 1-arachidonoyl glycerol bound to the cyclooxygenase channel of r513h murine cox-2

Structure:

Prostaglandin g/h synthase 2. Chain: a, b. Synonym: cyclooxygenase-2, cox-2, prostaglandin-endoperoxide synthase 2, prostaglandin h2 synthase 2, pgh synthase 2, pghs-2, phs ii, glucocorticoid-regulated inflammatory cyclooxygenase, gripghs, tis10 protein, macrophage activation-associated marker protein p71/73, pes- 2. Engineered: yes. Mutation: yes

Source:

Mus musculus. Mouse. Organism_taxid: 10090. Gene: ptgs2, cox-2, cox2, pghs-b, tis10. Expressed in: spodoptera frugiperda. Expression_system_taxid: 7108.

Resolution:

2.35Å R-factor: 0.164 R-free: 0.214

Authors:

A.J.Vecchio,M.G.Malkowski

Key ref:

A.J.Vecchio and M.G.Malkowski (2011). The structural basis of endocannabinoid oxygenation by cyclooxygenase-2. J Biol Chem, 286, 20736-20745. PubMed id: 21489986 DOI: 10.1074/jbc.M111.230367

Date:

26-Aug-10 Release date: 13-Apr-11

Protein chains

Q05769  (PGH2_MOUSE) -  Prostaglandin G/H synthase 2 from Mus musculus

Seq: 604 a.a.

Struc: 551 a.a.*

* PDB and UniProt seqs differ at 3 residue positions (black crosses)

Enzyme reactions

• Enzyme class: E.C.1.14.99.1 - prostaglandin-endoperoxide synthase.
• Reaction: (5Z,8Z,11Z,14Z)-eicosatetraenoate + AH2 + 2 O2 = prostaglandin H2 + A + H2O

(5Z,8Z,11Z,14Z)-eicosatetraenoate + AH2 + 2 × O2 = prostaglandin H2 (Bound ligand (Het Group name = COH) matches with 51.11% similarity) + + H2O

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

Added reference

DOI no: 10.1074/jbc.M111.230367

J Biol Chem 286:20736-20745 (2011)

PubMed id: 21489986

The structural basis of endocannabinoid oxygenation by cyclooxygenase-2.

A.J.Vecchio, M.G.Malkowski.

ABSTRACT

The cyclooxygenases (COX-1 and COX-2) oxygenate arachidonic acid (AA) in the committed step of prostaglandin biogenesis. Substitutions of I434V, H513R, and I523V constitute the only differences in residues lining the cyclooxygenase channel between COX-1 and COX-2. These changes create a hydrophobic pocket in COX-2, with Arg-513 located at the base of the pocket, which has been exploited in the design of COX-2-selective inhibitors. Previous studies have shown that COX-2, but not COX-1, can oxygenate endocannabinoid substrates, including 2-arachidonoyl glycerol (2-AG). To investigate the isoform-specific structural basis of endocannabinoid binding to COX-2, we determined the crystal structure of the 2-AG isomer 1-arachidonoyl glycerol (1-AG) in complex with wild type and R513H murine (mu) COX-2 to 2.2 and 2.35 Å, respectively, and R513H muCOX-2 in complex with AA to 2.45 Å resolution. The 2,3-dihydroxypropyl moiety of 1-AG binds near the opening of the cyclooxygenase channel in the space vacated by the movement of the Leu-531 side chain, validating our previous hypothesis implicating the flexibility of the Leu-531 side chain as a determinant for the ability of COX-2 to oxygenate endocannabinoid substrates. Functional analyses carried out to compliment our structural findings indicated that Y355F and R513H muCOX-2 constructs had no effect on the oxygenation of 1-AG and 2-AG, whereas substitutions that resulted in a shortened side chain for Leu-531 had only modest effects. Both AA and 1-AG bind to R513H muCOX-2 in conformations similar to those observed in the co-crystal structures of these substrates with wild type enzyme.