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首页> 外文期刊>Zoological Science >PP2A Type Phosphatases in Sea Urchin Eggs
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PP2A Type Phosphatases in Sea Urchin Eggs

机译：海胆鸡蛋中的PP2A型磷酸酶

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Four peaks of the p-nitrophenyl phosphate (pNPP) splitting activity were obtained by QAE-Toyopearl chromatography in the extract of sea urchin eggs from homogenate in 0.2M KCI solution containing 0.1% Triton X100 and so on. In 2 among these 4 peaks, pNPP splitting reactions were strongly inhibited by 10nM okadaic acid (OA) and calyculin A (CLA), potent and specific inhibitors of protein phosphatase 2A (PP2A). High sensitivities of pNPP splitting reaction to OA and CLA in these 2 peaks suggest that pNPP splitting results from the reaction catalyzed by PP2A, which reaction is expected to play roles in gene expression, signal transduction and cell movement such as cell division. OA sensitive pNPP splitting activities in these 2 peaks were eluted by EPLC gel permeation chromatography (Superdex 200HR) with approximate molecular mass of 160 kDa, corresponding to that of PP2A trimeric holoenzyme in mammalian cells. By immunoblot analyses with anti-human PP2A catalytic subunit antibody, an immunoreactive 36 kDa protein was found by SDS-PAGE in a peak of OA-sensitive pNPP splitting activity, obtained by QAE-Toyopearl chromatography. Sea urchin eggs have at least 2 PP2A like enzymes with similar molecular masses to that of mammalian PP2A and one of them contains human type catalytic subunit.
机译：通过QAE-Toyopearl色谱法在含有0.1％Triton X100的0.2M KCI溶液中从匀浆中提取海胆卵提取物中的磷酸对硝基苯酯（pNPP）裂解活性的四个峰。在这4个峰中的2个峰中，pNPP分裂反应被10nM冈田酸（OA）和calyculin A（CLA），蛋白磷酸酶2A的有效抑制剂和特异性抑制剂（PP2A）强烈抑制。在这两个峰中，pNPP裂解反应对OA和CLA的敏感性很高，这表明pNPP裂解是由PP2A催化的反应导致的，该反应有望在基因表达，信号转导和细胞运动（例如细胞分裂）中起作用。用EPLC凝胶渗透色谱法（Superdex 200HR）洗脱这两个峰中的OA敏感pNPP分裂活性，分子量约为160 kDa，相当于哺乳动物细胞中的PP2A三聚全酶。通过抗人PP2A催化亚基抗体的免疫印迹分析，通过SDS-PAGE在QAE-Toyopearl色谱法获得的OA敏感pNPP分裂活性峰值中发现了一种免疫反应性36 kDa蛋白。海胆卵至少含有2种PP2A类酶，其分子质量与哺乳动物PP2A相似，其中之一含有人型催化亚基。

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