Spodoptera frugiperda caspase-1, a novel insect death protease that cleaves the nuclear immunophilin FKBP46, is the target of the baculovirus antiapoptotic protein p35
Spodoptera frugiperda caspase-1, a novel insect death protease that cleaves the nuclear immunophilin FKBP46, is the target of the baculovirus antiapoptotic protein p35
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DOI：
10.1074/jbc.272.3.1421
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发表时间：
1997-01-17
影响因子：
4.8
通讯作者：
Alnemri, ES
中科院分区：
文献类型：
--
作者：
Ahmad, M;Srinivasula, SM;Alnemri, ES
利用最近用于克隆人Mch2的简并引物依赖的聚合酶链式反应方法，我们已经鉴定并克隆了昆虫夜蛾杆状病毒抗凋亡蛋白p35的靶标。该蛋白命名为SF caspase-1，属于caspase家族，在序列和比活性上与人Mch3和CPP32高度相关。Sf-caspase-1的酶原全长299个氨基酸，可在大肠杆菌中进行自催化反应，形成具有活性的酶异源复合体。Asp-28、Asp-184和Asp-195发生自动处理，以产生大的p19/p18和小的p12亚基。SF caspase-1能够诱导Sf9细胞的凋亡，并能够将p35切割成与p35缺失突变杆状病毒感染的Sf9细胞的提取物类似的大小片段。Sf caspase-1活性被p35有效抑制，提示它是该抗凋亡蛋白的一个重要靶点。最后，Sf9核免疫亲和素FKBP46被确定为SF caspase-1的死亡相关底物。
Employing the degenerate primer-dependent polymerase chain reaction approach used recently to clone human Mch2, we have identified and cloned the insect Spodoptera frugiperda target of the baculovirus antiapoptotic protein p35. This protein named Sf caspase-1 belongs to the family of caspases and is highly related to human Mch3 and CPP32 in sequence and specific activity. The proenzyme of Sf caspase-1 is 299 amino acids in length and can undergo autocatalytic processing in Escherichia coli to an active enzyme heterocomplex. Autoprocessing occurs at Asp-28, Asp-184, and Asp-195 to generate the large p19/p18 and small p12 subunits. Sf caspase-1 is able to induce apoptosis in Sf9 cells and is capable of cleaving p35 to similar sized fragments as observed with extracts from p35 null mutant baculovirus-infected Sf9 cells. Sf caspase-1 activity is potently inhibited by p35, suggesting that it is an important target of this antiapoptotic protein. Finally, the Sf9 nuclear immunophilin FKBP46 was identified as a death-associated substrate for Sf caspase-1.