A lung type prostaglandin F synthase is expressed in bovine liver: cDNA sequence and expression in E. coli.
A lung type prostaglandin F synthase is expressed in bovine liver: cDNA sequence and expression in E. coli.
复制标题
肺型前列腺素 F 合酶在牛肝中表达：cDNA 序列和在大肠杆菌中的表达。
DOI：
10.1016/s0006-291x(05)80323-7
复制
发表时间：
1992
影响因子：
3.1
通讯作者：
O. Hayaishi
中科院分区：
文献类型：
--
作者：
W. Kuchinke;O. Barski;K. Watanabe;O. Hayaishi
以牛肺前列腺素F合成酶(EC 1.1.1.2)的cDNA为探针，从牛肝cDNA文库中分离到一个与该探针仅有11个核苷酸差异的克隆。相应的蛋白质包含三个氨基酸替换，包括一个亮氨酸残基，它在所有的醛酮还原酶中都是保守的。将肝脏基因插入原核表达载体pUC19中，在大肠杆菌中表达重组肝酶。纯化的肝酶能还原前列腺素H_2、前列腺素D_2和各种羰基化合物。结合对前列腺素H_2的高相对活性和前列腺素D_2的高Km值，确定该酶为肺型前列腺素F合酶。但肝脏酶与NADPH的结合常数是肺前列腺素F合成酶的3.5倍。
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 inE.coli. The purified liver enzyme reduced prostaglandin H2as well as prostaglandin D2and various carbonyl compounds. The high relative activity against prostaglandin H2in combination with a high Km value for prostaglandin D2identified 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.
前列腺素 D2 经肝脏转化为新的前列腺素类化合物 9 α,11 β-前列腺素 F2，可抑制血小板聚集并收缩血管。
DOI：
--
发表时间：
1985
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Pugliese,G;Spokas,EG;Marcinkiewicz,E;Wong,PY
通讯作者：
Wong,PY