大鼠嗜碱性白血病细胞中的12-脂氧合酶：与5-脂氧合酶的分离及氢过氧脂肪酸对其温度依赖性失活作用
12-Lipoxygenase from rat basophilic leukemia cells: separation from 5-lipoxygenase and temperature-dependent inactivation by hydroperoxy fatty acid.
作者信息
van der Donk E M, Verhagen J, Veldink G A, Vliegenthart J F
机构信息
Department of Bio-Organic Chemistry, Utrecht University, The Netherlands.
出版信息
Biochim Biophys Acta. 1991 Jan 28;1081(2):135-40. doi: 10.1016/0005-2760(91)90018-d.
12-Lipoxygenase and 5-lipoxygenase from rat basophilic leukemia cells were separated by protein-HPLC in a single step. Upon incubation in the presence of Ca2+, 12-lipoxygenase converted arachidonic acid into 12(S)-hydroxyeicosatetraenoic acid and linoleic acid into 13(S)-hydro(pero)xyoctadecadienoic acid. The reaction products were analyzed by reversed-phase and chiral straight-phase HPLC with ultraviolet-detection. Using the cytosolic fraction of rat basophilic leukemia cells, optimal 12-lipoxygenase activity was observed at 10 degrees C. At 37 degrees C 12-lipoxygenase was very rapidly inactivated by its own product, hydroperoxy fatty acid, at low concentrations (10-100 nM).
大鼠嗜碱性白血病细胞中的12-脂氧合酶和5-脂氧合酶通过蛋白质高效液相色谱法一步分离。在Ca2+存在下孵育时，12-脂氧合酶将花生四烯酸转化为12(S)-羟基二十碳四烯酸，并将亚油酸转化为13(S)-氢（过氧）十八碳二烯酸。通过反相和手性正相高效液相色谱法结合紫外检测对反应产物进行分析。使用大鼠嗜碱性白血病细胞的胞质部分，在10℃观察到最佳的12-脂氧合酶活性。在37℃时，12-脂氧合酶在低浓度（10-100 nM）的自身产物氢过氧脂肪酸作用下非常迅速地失活。
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