描述两个交替的拼接形式的磷脂酶D1。纯化酶的激活磷脂酰肌醇4,5-bisphosphate ADP-ribosylation因素,ρ家庭单体的GTP-binding蛋白质和蛋白激酶C-alpha。
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哈蒙德SM, Jenco JM,中岛美嘉年代,卡德瓦拉德K,顾Q,库克,Nozawa Y, Prestwich GD, Frohman妈,莫里斯AJ
描述两个交替的拼接形式的磷脂酶D1。纯化酶的激活磷脂酰肌醇4,5-bisphosphate ADP-ribosylation因素,ρ家庭单体的GTP-binding蛋白质和蛋白激酶C-alpha。
生物化学杂志。1997年2月7日,272 (6):3860 - 8。
- PubMed ID
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9013646 (在PubMed]
- 文摘
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我们之前报道的cDNA克隆编码人类phosphatidylcholine-specific磷脂酶D1 (PLD1),一个ADP-ribosylation因素(ARF)激活phosphatidylcholine-specific磷脂酶D(哈蒙德,s M。Tsung年代。Autschuller, Y。,Rudge s。,玫瑰,K。Engebrecht, J。莫里斯,a·J。,Frohman m a(1995)生物。270年化学,29640 - 29643)。我们已经确定了一个进化保存短接头的变体PLD1缺乏38氨基酸(残留585 - 624),来自监管的另一个外显子剪接。两种PLD1 (PLD1a和1 b)一直在表达Sf9细胞利用杆状病毒载体和提取和immunoaffinity色谱纯化的同质性的洗衣粉。PLD1a和1 b非常相似的属性。PLD1a和Mg2 + 1 b活动相关但不自由钙离子浓度的变化。磷脂酰肌醇4,5-bisphosphate和磷脂酰肌醇3、4、5-trisphosphate激活PLD1a和1 b,但一系列其他酸性磷脂是无效的。 PLD1a and 1b are highly responsive to activation by GTP-gammaS-liganded ADP-ribosylation factor-1 (ARF-1) and can also be activated to a lesser extent by three purified RHO family monomeric GTP-binding proteins, RHO A, RAC-1, and CDC42. Activation of PLD1a and 1b by the RHO family monomeric GTP-binding proteins is GTP-dependent and synergistic with ARF-1. Purified protein kinase C-alpha activates PLD1a and 1b in a manner that is stimulated by phorbol esters and does not require ATP. Activation of PLD1a and 1b by protein kinase C-alpha is synergistic with ARF and with the RHO family monomeric GTP-binding proteins, suggesting that these three classes of regulators interact with different sites on the enzyme.