Synthesis of prostaglandin F ethanolamide by prostaglandin F synthase and identification of Bimatoprost as a potent inhibitor of the enzyme: new enzyme assay method using LC/ESI/MS
Synthesis of prostaglandin F ethanolamide by prostaglandin F synthase and identification of Bimatoprost as a potent inhibitor of the enzyme: new enzyme assay method using LC/ESI/MS
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DOI：
10.1016/j.abb.2004.02.009
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发表时间：
2004-04-15
影响因子：
3.9
通讯作者：
Watanabe, K
中科院分区：
文献类型：
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作者：
Koda, N;Tsutsui, Y;Watanabe, K
前列腺素 (PG) D-2 乙醇酰胺（前列腺酰胺 D-2）被 PGF 合酶还原为 9α, 11β-PGF(2) 乙醇酰胺（9α, 11β-前列腺酰胺 F-2），该酶还催化 PGH(2) 和 PGD(2) 分别还原为 PGF(2α) 和 9α11β-PGF(2)。这些酶的活性通过液相色谱-电喷雾电离-质谱（LC/ESI/MS）技术来测量，该技术可以同时检测底物和所有产物。 PGF(2α)、9α,11β-PGF(2)、PGD(2)、PGH(2)、9α,11β-前列腺酰胺F-2和前列腺酰胺D-2在TSKgel ODS 80Ts柱上分离，通过电喷雾电离，并在负模式下检测。 m/z 353 ([M-H](-))、353 ([M-H](-))、351 ([M-H](-)) 333 ([M-H-H2O](-)) 456 ([M+59](-)) 和 m/z 358 ([M-37](-)) 的选择离子监测 (SIM) 用于定量分别为 PGF(2α)、9α,11β-PGF(2)、PGD(2)、PGH(2)、9α,11β-前列腺酰胺 F-2 和前列腺酰胺 D-2。 PGF(2α)和9α,11β-PGF(2)的检测限为0.01 pmol，PGH(2)和PGD(2)的检测限为0.1 pmol；前列腺酰胺 D-2 和 9α,11β-前列腺酰胺 F-2 分别为 0.5 和 0.03 pmol。用于测量 PGF 合酶活性的 LC/ESI/MS 技术显示出比其他方法更高的灵敏度。使用这种方法，我们发现比马前列素（17-苯基-三去甲 PGF(2α) 的乙酰胺类似物和抗青光眼剂）在低浓度使用时会抑制 PGF 合酶的所有三种还原酶活性。这些结果表明，比马前列素除了具有前列腺酰胺样活性外，还可以作为有效的 PGF 合酶抑制剂。 (C) 2004 Elsevier Inc. 保留所有权利。
Prostaglandin (PG) D-2 ethanolamide (prostamide D-2) was reduced to 9alpha, 11beta-PGF(2) ethanolamide (9alpha, 11beta-prostamide F-2) by PGF synthase, which also catalyzes the reduction of PGH(2) and PGD(2) to PGF(2alpha) and 9alpha11beta-PGF(2), respectively. These enzyme activities were measured by a new method, the liquid chromatographic-electrospray ionization-mass spectrometry (LC/ESI/MS) technique, which could simultaneously detect the substrate and all products. PGF(2alpha), 9alpha,l lbeta-PGF(2), PGD(2), PGH(2), 9alpha,11beta-prostamide F-2, and prostamide D-2 were separated on a TSKgel ODS 80Ts column, ionized by electrospray, and detected in the negative mode. Selected ion monitoring (SIM) of m/z 353 ([M-H](-)), 353 ([M-H](-)), 351 ([M-H](-)) 333 ([M-H-H2O](-)) 456 ([M+59](-)), and m/z 358 ([M-37](-)) was used for quantifying PGF(2alpha), 9alpha,11beta-PGF(2), PGD(2), PGH(2), 9alpha,11beta-prostamide F-2, and prostamide D-2, respectively. The detection limit for PGF(2alpha) and 9alpha,11beta-PGF(2) was 0.01 pmol, that for PGH(2) and PGD(2), 0.1 pmol; and that for prostamide D-2 and 9alpha,11beta-prostamide F-2, 0.5 and 0.03 pmol, respectively. The LC/ESI/MS technique for measuring PGF synthase activity showed higher sensitivity than other methods. Using this method, we found that Bimatoprost, the ethyl amide analog of 17-phenyl-trinor PGF(2alpha) and an anti-glaucoma agent, inhibited all three reductase activities of PGF synthase when used at a low concentration. These results suggest that Bimatoprost also behaves as a potent PGF synthase inhibitor in addition to having prostamide-like activity. (C) 2004 Elsevier Inc. All rights reserved.