On non-cyclooxygenase prostaglandin synthesis in the sea whip coral, Plexaura homomalla: an 8(R)-lipoxygenase pathway leads to formation of an alpha-ketol and a Racemic prostanoid.
Plexaura homomalla is a rich natural source of prostaglandins and recent evidence suggest the prostaglandin biosynthesis could occur through a lipoxygenase pathway. We have investigated the metabolism of arachidonic acid in homogenates and acetone powders of the fresh frozen coral. The biosynthesis of natural prostaglandins was not detected. However, we find a prominent 8(R)-lipoxygenase pathway leading to an alpha-ketol, characterized by high pressure liquid chromatography, gas chromatography-mass spectrometry, and NMR as 8-hydroxy, 9-keto-eicosa-5Z, 11Z, 14Z-trienoic acid, and a prostaglandin A-like cyclopentenone identified as 9-oxo-[8, 12-cis]-prosta-5Z, 10, 14Z-trienoic acid. These reactions appear analogous to the transformation of linolenic acid hydroperoxide by "isomerase" and "cyclase" of corn and flaxseed. From analysis of the absolute configurations of the coral products, and from additional stable isotope labeling experiments in H218O and D2O, we deduce that both compounds arise via conversion of 8(R)-HPETE to an 8(R), 9-allene oxide, 8R,9-oxido-eicosa-5Z, 9, 11Z, 14Z-tetraenoic acid. This unstable intermediate undergoes hydrolysis to form the alpha-ketol or cyclization to give the cyclopentenone. Significantly, we find that the prostaglandin-like product is a racemic mixture of cis side chain enantiomers, pointing to its nonenzymatic origin from the allene oxide. The alpha-ketol is formed with partial racemization and inversion of configuration, also compatible with formation in a nonenzymatic reaction. We conclude that the isomerase and cyclase reactions may merely reflect nonenzymatic breakdown of the enzymatically formed allene oxide. The origin of the endogenous (chiral) prostaglandins of the coral may involve an allene oxide intermediate, although the potential for formation of racemic products presents an interesting dilemma regarding its relationship to the natural pathway of biosynthesis.
Brash
A R
AR
Department of Pharmacology, Vanderbilt University, Nashville, Tennessee 37232.
Baertschi
S W
SW
Ingram
C D
CD
Harris
T M
TM
D000818:Animals; D001091:Arachidonate Lipoxygenases / Q000378:metabolism*; D016718:Arachidonic Acid; D001095:Arachidonic Acids / Q000378:metabolism; D002250:Carbon Radioisotopes; D002851:Chromatography, High Pressure Liquid; D003903:Deuterium; D017666:Deuterium Oxide; D007700:Kinetics; D008974:Mollusca / Q000201:enzymology*; D010103:Oxygen Isotopes; D011451:Prostaglandin-Endoperoxide Synthases; D011453:Prostaglandins / Q000096:biosynthesis*; D014867:Water
D001095:Arachidonic Acids; D002250:Carbon Radioisotopes; D010103:Oxygen Isotopes; D011453:Prostaglandins; D014867:Water; D016718:Arachidonic Acid; D003903:Deuterium; D001091:Arachidonate Lipoxygenases; C054192:arachidonate 8-lipoxygenase; D011451:Prostaglandin-Endoperoxide Synthases; D017666:Deuterium Oxide