Interaction of 5-lipoxygenase with cellular proteins
Interaction of 5-lipoxygenase with cellular proteins
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DOI：
10.1073/pnas.96.5.1881
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发表时间：
1999-03-02
影响因子：
11.1
通讯作者：
Rådmark, O
中科院分区：
文献类型：
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作者：
Provost, P;Samuelsson, B;Rådmark, O
5-脂氧合酶(5LO)在细胞白三烯合成中起着关键作用。为了鉴定与人5LO相互作用的蛋白质，我们使用双杂交方法筛选了一个人肺cDNA文库。从总共1.5×10(7)个酵母转化子中，分离出代表三种不同蛋白质的9个独立克隆，并发现它们与5LO特异相互作用。四个1.7-1.8kb的克隆代表了一个16 kDa的蛋白，命名为coactosin-like蛋白，因为它与coactosin有显著的同源性，coactosin是一种与盘基网眼菌中的肌动蛋白相关的蛋白。因此，类Coactosin蛋白可能在5LO和细胞骨架之间提供了联系。另外两个1.5kb的酵母克隆编码转化生长因子型β受体I相关蛋白1部分基因。转化生长因子β受体I相关蛋白1最近被报道与转化生长因子β受体I的激活形式有关，并可能参与转化生长因子β诱导HL-60和Mono Mac 6细胞中5LO表达和活性的上调。最后，3个全长2.1kb的克隆含有一个与秀丽线虫假定的解旋酶K12H4.8高度同源的人类蛋白的部分cDNA，被命名为Delta K12H4.8同源物。对预测的氨基酸序列的分析表明，存在一个RNaseIII基序和一个双链RNA结合域，表明这是一种核起源的蛋白质。这些5LO相互作用蛋白的鉴定为研究5LO的细胞功能提供了新的途径。
5-Lipoxygenase (5LO) plays a pivotal role in cellular leukotriene synthesis. To identify proteins interacting with human 5LO, we used a two-hybrid approach to screen a human lung cDNA library. From a total of 1.5 x 10(7) yeast transformants, nine independent clones representing three different proteins were isolated and found to specifically interact with 5LO. Four 1.7- to 1.8-kb clones represented a 16-kDa protein named coactosin-like protein for its significant homology with coactosin, a protein found to be associated with actin in Dictyostelium discoideum. Coactosin-like protein thus may provide a link between 5LO and the cytoskeleton. Two other yeast clones of 1.5 kb encoded transforming growth factor (TGF) type beta receptor-I-associated protein 1 partial cDNA. TGF type beta receptor-I-associated protein 1 recently has been reported to associate with the activated form of the TGF beta receptor I and may be involved in the TGF beta-induced up-regulation of 5LO expression and activity observed in HL-60 and Mono Mac 6 cells. Finally, three identical 2.1-kb clones contained the partial cDNA of a human protein with high homology to a hypothetical helicase K12H4.8 from Caenorhabditis elegans and consequently was named Delta K12H4.8 homologue. Analysis of the predicted amino acid sequence revealed the presence of a RNase III motif and a double-stranded RNA binding domain, indicative of a protein of nuclear origin. The identification of these 5LO-interacting proteins provides additional approaches to studies of the cellular functions of 5LO.