Prostaglandin production from arachidonic acid and evidence for a 9,11-endoperoxide prostaglandin H2 reductase in Leishmania

• Published in: International journal for parasitology Vol. 32; no. 14; pp. 1693 - 1700
• Main Authors: KABUTUTU, Zakayi, MARTIN, Samuel K, KILUNGA KUBATA, Bruno, NOZAKI, Tomoyoshi, KAWAZU, Shin-Ichiro, OKADA, Tetsuya, MUNDAY, Craig Joe, DUSZENKO, Michael, LAZARUS, Michael, THUITA, Lucy W, URADE, Yoshihiro
• Format: Journal Article
• Language: English
• Published: Oxford Elsevier Science 19.12.2002
• Subjects: Animals; Arachidonic Acid - metabolism; Base Sequence; Biochemistry. Physiology. Immunology. Molecular biology; Biological and medical sciences; Blotting, Western; DNA, Complementary - genetics; DNA, Protozoan - genetics; Fundamental and applied biological sciences. Psychology; Hydroxyprostaglandin Dehydrogenases - genetics; Hydroxyprostaglandin Dehydrogenases - metabolism; Leishmania - classification; Leishmania - enzymology; Leishmania - metabolism; Mice; Mice, Inbred BALB C; Molecular Sequence Data; Polymerase Chain Reaction - methods; Prostaglandin H2; Prostaglandins - biosynthesis; Prostaglandins H - metabolism; Protozoa; Recombinant Proteins - metabolism; Trypanosoma brucei brucei - enzymology; Host parasite relation; Protozoal disease; Prostaglandin-F synthase; Kinetoplastid; Leishmania donovani; Biosynthesis; Arachidonic acid; Enzymatic activity; Trypanosoma brucei; Kinetoplastida; Protozoa; Prostaglandin; Enzyme; Arachidonic acid derivatives; Parasite; Arachidonic acid metabolism; Parasitosis; Leishmaniasis; Metabolism; Prostaglandin F2α synthase; Prostaglandin production; Infection; Trypanosomatid; Leishmania; Leishmania tropica; Oxidoreductases; Leishmania major
• ISSN: 0020-7519; 1879-0135
• DOI: 10.1016/s0020-7519(02)00160-1

Lysates of Leishmania promastigotes can metabolise arachidonic acid to prostaglandins. Prostaglandin production was heat sensitive and not inhibited by aspirin or indomethacin. We cloned and sequenced the cDNA of Leishmania major, Leishmania donovani, and Leishmania tropica prostaglandin F(2alpha) synthase, and overexpressed their respective 34-kDa recombinant proteins that catalyse the reduction of 9,11-endoperoxide PGH(2) to PGF(2alpha). Database search and sequence alignment alignment showed that L. major prostaglandin F(2alpha) synthase exhibits 61, 99.3, and 99.3% identity with Trypanosoma brucei, L. donovani, and L. tropica prostaglandin F(2alpha) synthase, respectively. Using polymerase chain reaction amplification, Western blotting, and immunofluorescence, we have demonstrated that prostaglandin F(2alpha) synthase protein and gene are present in Old World and absent in New World Leishmania, and that this protein is localised to the promastigote cytosol.