Prostaglandin production from arachidonic acid and evidence for a 9,11-endoperoxide prostaglandin H2 reductase in Leishmania.
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 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.
Kabututu
Zakayi
Z
Department of Molecular Behavioral Biology, Osaka Bioscience Institute, Suita, Osaka 565-0874, Japan.
Martin
Samuel K
SK
Nozaki
Tomoyoshi
T
Kawazu
Shin-ichiro
S
Okada
Tetsuya
T
Munday
Craig Joe
CJ
Duszenko
Michael
M
Lazarus
Michael
M
Thuita
Lucy W
LW
Urade
Yoshihiro
Y
Kubata
Bruno Kilunga
BK
D000818:Animals; D016718:Arachidonic Acid / Q000378:metabolism*; D001483:Base Sequence; D015153:Blotting, Western; D015237:Dinoprost / Q000096:biosynthesis*; D005455:Fluorescent Antibody Technique; D006911:Hydroxyprostaglandin Dehydrogenases / Q000235:genetics / Q000378:metabolism*; D007891:Leishmania / Q000378:metabolism*; D007893:Leishmania donovani / Q000235:genetics; D007895:Leishmania tropica / Q000235:genetics; D008969:Molecular Sequence Data; D016415:Sequence Alignment; D017385:Sequence Homology; D014346:Trypanosoma brucei brucei / Q000235:genetics
D016718:Arachidonic Acid; D015237:Dinoprost; D006911:Hydroxyprostaglandin Dehydrogenases; C031051:prostaglandin-F synthase