title: The origin of 15R-prostaglandins in the Caribbean coral Plexaura homomalla: molecular cloning and expression of a novel cyclooxygenase.
abstract: The highest concentrations of prostaglandins in nature are found in the Caribbean gorgonian Plexaura homomalla. Depending on its geographical location, this coral contains prostaglandins with typical mammalian stereochemistry (15S-hydroxy) or the unusual 15R-prostaglandins. Their metabolic origin has remained the subject of mechanistic speculations for three decades. Here, we report the structure of a type of cyclooxygenase (COX) that catalyzes transformation of arachidonic acid into 15R-prostaglandins. Using a homology-based reverse transcriptase--PCR strategy, we cloned a cDNA corresponding to a COX protein from the R variety of P. homomalla. The deduced peptide sequence shows 80% identity with the 15S-specific coral COX from the Arctic soft coral Gersemia fruticosa and approximately 50% identity to mammalian COX-1 and COX-2. The predicted tertiary structure shows high homology with mammalian COX isozymes having all of the characteristic structural units and the amino acid residues important in catalysis. Some structural differences are apparent around the peroxidase active site, in the membrane-binding domain, and in the pattern of glycosylation. When expressed in Sf9 cells, the P. homomalla enzyme forms a 15R-prostaglandin endoperoxide together with 11R-hydroxyeicosatetraenoic acid and 15R-hydroxyeicosatetraenoic acid as by-products. The endoperoxide gives rise to 15R-prostaglandins and 12R-hydroxyheptadecatrienoic acid, identified by comparison to authentic standards. Evaluation of the structural differences of this 15R-COX isozyme should provide new insights into the substrate binding and stereospecificity of the dioxygenation reaction of arachidonic acid in the cyclooxygenase active site.
authors: Department of Bioorganic Chemistry, Institute of Chemistry at Tallinn Technical University, Akadeemia tee 15, Tallinn 12618, Estonia.
authors: K
authors: K
authors: Valmsen
authors: I
authors: I
authors: Järving
authors: W E
authors: WE
authors: Boeglin
authors: K
authors: K
authors: Varvas
authors: R
authors: R
authors: Koljak
authors: T
authors: T
authors: Pehk
authors: A R
authors: AR
authors: Brash
authors: N
authors: N
authors: Samel
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D003001:Cloning, Molecular; D003063:Cnidaria / Q000201:enzymology* / Q000235:genetics*; D015232:Dinoprostone / Q000031:analogs & derivatives / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D011451:Prostaglandin-Endoperoxide Synthases / Q000235:genetics* / Q000378:metabolism; D011453:Prostaglandins / Q000235:genetics* / Q000378:metabolism; D011454:Prostaglandins A / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D013237:Stereoisomerism
chemical_list: D011453:Prostaglandins; D011454:Prostaglandins A; C041324:prostaglandin E2 methyl ester; D011451:Prostaglandin-Endoperoxide Synthases; D015232:Dinoprostone