cDNA cloning, expression and characterization of human prostaglandin F synthase.
A cDNA clone of prostaglandin F synthase (PGFS) was isolated from human lung by using cDNA of bovine lung-type PGFS as a probe and its protein expressed in Escherichia coli was purified to apparent homogeneity. The human PGFS catalyzed the reduction of prostaglandin (PG) D2, PGH2 and phenanthrenequinone (PQ), and the oxidation of 9alpha,11beta-PGF2 to PGD2. The kcat/Km values for PGD2 and 9alpha,11beta-PGF2 were 21000 and 1800 min(-1) mM(-1), respectively, indicating that the catalytic efficiency for PGD2 and 9alpha,11beta-PGF2 was the highest among the various substrates, except for PQ. The PGFS activity in the cytosol of human lung was completely absorbed with antihuman PGFS antiserum. Moreover, mRNA of PGFS was expressed in peripheral blood lymphocytes and the expression in lymphocytes was markedly suppressed by the T cell mitogen concanavalin A. These results support the notion that human PGFS plays an important role in the pathogenesis of allergic diseases such as asthma.
Suzuki-Yamamoto
T
T
Department of Anatomy and Cell Biology, School of Medicine, The University of Tokushima, Kuramoto, Japan.
Nishizawa
M
M
Fukui
M
M
Okuda-Ashitaka
E
E
Nakajima
T
T
Ito
S
S
Watanabe
K
K
D000595:Amino Acid Sequence; D001483:Base Sequence; D015153:Blotting, Western; D003001:Cloning, Molecular; D046508:Culture Techniques; D018076:DNA, Complementary / Q000378:metabolism; D004926:Escherichia coli / Q000378:metabolism; D015723:Gene Library; D006801:Humans; D006911:Hydroxyprostaglandin Dehydrogenases / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D007700:Kinetics; D008168:Lung / Q000201:enzymology*; D008214:Lymphocytes / Q000201:enzymology; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction
D018076:DNA, Complementary; D011994:Recombinant Proteins; D006911:Hydroxyprostaglandin Dehydrogenases; C031051:prostaglandin-F synthase