9 alpha,11 beta-prostaglandin F2 formation in various bovine tissues. Different isozymes of prostaglandin D2 11-ketoreductase, contribution of prostaglandin F synthetase and its cellular localization.
9 alpha,11 beta-prostaglandin F2 formation in various bovine tissues. Different isozymes of prostaglandin D2 11-ketoreductase, contribution of prostaglandin F synthetase and its cellular localization.
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9 α,11 β-前列腺素 F2 在各种牛组织中形成。
DOI：
10.1016/s0021-9258(19)38503-5
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发表时间：
1990
期刊：
影响因子：
--
通讯作者：
O. Hayaishi
中科院分区：
文献类型：
--
作者：
Y. Urade;Kikuko Watanabe;N. Eguchi;Yutaka Fujii;O. Hayaishi
在 NADPH 生成系统存在的情况下，在各种牛组织的 100,000 x g 上清液中，通过前列腺素 D2 的 11-酮还原酶形成 9 α,11 β-前列腺素 F2。肝脏（51.09 nmol/h/mg 蛋白质）、肺（24.99）和脾（14.20）中还原酶活性较高；心脏和胰腺中等（3.09-3.61）；胃、肠、结肠、肾、子宫、肾上腺、胸腺虚弱（0.11-2.63）；在大脑、视网膜、颈动脉和血液中检测不到（小于 0.10）。在所有组织中均未检测到前列腺素 D2 形成前列腺素 F2 α。在用前列腺素F合成酶特异性多克隆抗体进行免疫滴定分析时，肺和脾中的还原酶活性显示出与纯化合成酶相同的滴定曲线，并且在抗体过量的情况下降低至初始活性的15%以下。这两种组织中的前列腺素 F 合成酶免疫反应蛋白显示出与金黄色葡萄球菌 V8 蛋白酶部分消化后纯化酶相同的肽指纹。该抗体与肝脏中的还原酶发生部分交叉反应（大约与合成酶发生交叉反应的 20%），但与其他组织中的还原酶不发生交叉反应。前列腺素 D2 还原酶活性的 Km 值在肺和脾中与纯化的前列腺素 F 合成酶 (120 µM) 相同，但在肝脏 (6 µM)、心脏和胰腺 (15 µM) 中不同。前列腺素 F 合成酶在肺和脾中的主要分布通过放射免疫测定（分别为 2.8 和 1.0 微克/毫克蛋白质）和 Northern 印迹分析得到证实。在免疫过氧化物酶染色中，该酶定位于肺泡间质细胞和肺中的无纤毛上皮细胞、脾中的组织细胞和/或树突状细胞以及肾和肾上腺皮质中的一些间质细胞。
9 alpha,11 beta-prostaglandin F2 was formed from prostaglandin D2 by its 11-ketoreductases in 100,000 x g supernatants of various bovine tissues in the presence of an NADPH-generating system. The reductase activities were high in liver (51.09 nmol/h/mg of protein), lung (24.99), and spleen (14.20); moderate in heart and pancreas (3.09-3.61); weak in stomach, intestine, colon, kidney, uterus, adrenal gland, and thymus (0.11-2.63); and undetectable in brain, retina, carotid artery, and blood (less than 0.10). No formation of prostaglandin F2 alpha from prostaglandin D2 was detected in all tissues. In immunotitration analyses with a polyclonal antibody specific for prostaglandin F synthetase, the reductase activities in lung and spleen showed identical titration curves to that of the purified synthetase and decreased to less than 15% of the initial activity under the condition of antibody excess. Prostaglandin F synthetase-immunoreactive protein in these two tissues showed peptide fingerprints identical to that of the purified enzyme after partial digestion with Staphylococcus aureus V8 protease. The antibody was partially cross-reactive to the reductase in liver (about 20% of that to the synthetase) but not to the reductase(s) in other tissues. The Km value for prostaglandin D2 of the reductase activity was the same in lung and spleen as that of the purified prostaglandin F synthetase (120 microM) but differed in liver (6 microM), heart, and pancreas (15 microM). The predominant distribution of prostaglandin F synthetase in lung and spleen was confirmed by radioimmunoassay (2.8 and 1.0 micrograms/mg protein, respectively) and Northern blot analyses. In immunoperoxidase staining, this enzyme was localized in alveolar interstitial cells and nonciliated epithelial cells in lung, histiocytes and/or dendritic cells in spleen, and a few interstitial cells in kidney and adrenal cortex.