The basis of prostaglandin synthesis in coral: molecular cloning and expression of a cyclooxygenase from the Arctic soft coral Gersemia fruticosa.

Koljak R, Järving I, Kurg R, Boeglin WE, Varvas K, Valmsen K, Ustav M, Brash AR, Samel N
J Biol Chem. 2001 276 (10): 7033-40

PMID: 11085996 · DOI:10.1074/jbc.M009803200

In vertebrates, the synthesis of prostaglandin hormones is catalyzed by cyclooxygenase (COX)-1, a constitutively expressed enzyme with physiological functions, and COX-2, induced in inflammation and cancer. Prostaglandins have been detected in high concentrations in certain corals, and previous evidence suggested their biosynthesis through a lipoxygenase-allene oxide pathway. Here we describe the discovery of an ancestor of cyclooxygenases that is responsible for prostaglandin biosynthesis in coral. Using a homology-based polymerase chain reaction cloning strategy, the cDNA encoding a polypeptide with approximately 50% amino acid identity to both mammalian COX-1 and COX-2 was cloned and sequenced from the Arctic soft coral Gersemia fruticosa. Nearly all the amino acids essential for substrate binding and catalysis as determined in the mammalian enzymes are represented in coral COX: the arachidonate-binding Arg(120) and Tyr(355) are present, as are the heme-coordinating His(207) and His(388); the catalytic Tyr(385); and the target of aspirin attack, Ser(530). A key amino acid that determines the sensitivity to selective COX-2 inhibitors (Ile(523) in COX-1 and Val(523) in COX-2) is present in coral COX as isoleucine. The conserved Glu(524), implicated in the binding of certain COX inhibitors, is represented as alanine. Expression of the G. fruticosa cDNA afforded a functional cyclooxygenase that converted exogenous arachidonic acid to prostaglandins. The biosynthesis was inhibited by indomethacin, whereas the selective COX-2 inhibitor nimesulide was ineffective. We conclude that the cyclooxygenase occurs widely in the animal kingdom and that vertebrate COX-1 and COX-2 are evolutionary derivatives of the invertebrate precursor.

MeSH Terms (32)

Alanine Amino Acid Sequence Animals Arginine Blotting, Northern Chromatography, Thin Layer Cloning, Molecular Cnidaria COS Cells Cyclooxygenase 1 Cyclooxygenase 2 DNA, Complementary HeLa Cells Histidine Humans Isoenzymes Isoleucine Membrane Proteins Microscopy, Fluorescence Models, Genetic Molecular Sequence Data Phylogeny Plasmids Polymerase Chain Reaction Prostaglandin-Endoperoxide Synthases Prostaglandins Protein Binding RNA, Messenger Sequence Analysis, DNA Sequence Homology, Amino Acid Serine Tyrosine

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