A lung type prostaglandin F synthase is expressed in bovine liver: cDNA sequence and expression in E. coli.
Using the cDNA of bovine lung prostaglandin F synthase (EC 1.1.1.2) as a probe, we isolated a clone from a bovine liver cDNA library which differed in only eleven nucleotides from the probe. The corresponding protein contained three amino acid substitutions, including a leucine residue which is conserved throughout all aldo-keto reductases. We inserted the liver cDNA into expression vector pUC19 and expressed the recombinant liver enzyme in E.coli. The purified liver enzyme reduced prostaglandin H2 as well as prostaglandin D2 and various carbonyl compounds. The high relative activity against prostaglandin H2 in combination with a high Km value for prostaglandin D2 identified this liver enzyme as a lung type prostaglandin F synthase. However, the binding constant for NADPH of the liver enzyme was 3.5 fold higher than that of lung prostaglandin F synthase.
Kuchinke
W
W
Osaka Bioscience Institute, Japan.
Barski
O
O
Watanabe
K
K
Hayaishi
O
O
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002417:Cattle; D004277:DNA, Single-Stranded; D015237:Dinoprost / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics; D015723:Gene Library; D006911:Hydroxyprostaglandin Dehydrogenases / Q000145:classification / Q000235:genetics*; D008099:Liver / Q000201:enzymology*; D008168:Lung / Q000201:enzymology; D008969:Molecular Sequence Data; D015230:Prostaglandin D2 / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism; D012689:Sequence Homology, Nucleic Acid; D013379:Substrate Specificity
D004277:DNA, Single-Stranded; D011994:Recombinant Proteins; D015237:Dinoprost; D006911:Hydroxyprostaglandin Dehydrogenases; C031051:prostaglandin-F synthase; D015230:Prostaglandin D2