Cleavage of lamin A by Mch2 alpha but not CPP32: multiple interleukin 1 beta-converting enzyme-related proteases with distinct substrate recognition properties are active in apoptosis.
Cleavage of lamin A by Mch2 alpha but not CPP32: multiple interleukin 1 beta-converting enzyme-related proteases with distinct substrate recognition properties are active in apoptosis.
复制标题
DOI：
10.1073/pnas.93.16.8395
复制
发表时间：
1996-08
影响因子：
11.1
通讯作者：
A. Takahashi;A. Takahashi;E. Alnemri;Y. Lazebnik;Y. Lazebnik;T. Fernandes‐Alnemri;G. Litwack;R. Moir;R. Goldman;G. Poirier;S. Kaufmann;W. Earnshaw;W. Earnshaw
中科院分区：
文献类型：
--
作者：
A. Takahashi;A. Takahashi;E. Alnemri;Y. Lazebnik;Y. Lazebnik;T. Fernandes‐Alnemri;G. Litwack;R. Moir;R. Goldman;G. Poirier;S. Kaufmann;W. Earnshaw;W. Earnshaw
尽管与白介素1β转换酶(ICE)相关的蛋白水解酶在细胞凋亡中起重要作用，但目前尚不清楚参与其中的酶的数量、底物的特异性及其在细胞凋亡的生化和形态变化中的具体作用。利用克隆的重组ICE相关蛋白(IRPS)和无细胞的细胞凋亡模型系统(S/M提取物)解决了这些问题。首先，我们比较了两种重组人IRPS，CPP32和Mch2α的底物特异性。这两种酶都能裂解多聚(ADP-核糖)聚合酶，尽管效率不同。Mch2α还在一个保守的VEID处切割重组和核层蛋白A，减少位于盘状杆状结构域中间的NG序列，产生一个与S/M提取物和凋亡细胞中观察到的层蛋白A片段难以区分的片段。Mch2α和S/M提取物对Lamin A的裂解被毫摩尔浓度的锌离子抑制，而对CPP32和S/M提取物切割聚腺苷二磷酸核糖聚合酶的影响最小。我们还发现，衍生活性ICE大亚基的N-(乙酰酪氨酸-N-β-生物素赖氨基)天冬氨酸[(2，6-二甲基苯甲酰氧基)甲基酮]甲酮在S/M提取物中可以亲和标记多达5个活性IRP。综上所述，这些观察表明，核蛋白在凋亡中的处理涉及多个IRP，它们对与凋亡相关的底物具有不同的偏好。
Although proteases related to the interleukin 1 beta-converting enzyme (ICE) are known to be essential for apoptotic execution, the number of enzymes involved, their substrate specificities, and their specific roles in the characteristic biochemical and morphological changes of apoptosis are currently unknown. These questions were addressed using cloned recombinant ICE-related proteases (IRPs) and a cell-free model system for apoptosis (S/M extracts). First, we compared the substrate specificities of two recombinant human IRPs, CPP32 and Mch2 alpha. Both enzymes cleaved poly-(ADP-ribose) polymerase, albeit with different efficiencies. Mch2 alpha also cleaved recombinant and nuclear lamin A at a conserved VEID decreases NG sequence located in the middle of the coiled-coil rod domain, producing a fragment that was indistinguishable from the lamin A fragment observed in S/M extracts and in apoptotic cells. In contrast, CPP32 did not cleave lamin A. The cleavage of lamin A by Mch2 alpha and by S/M extracts was inhibited by millimolar concentrations of Zn2+, which had a minimal effect on cleavage of poly (ADP-ribose) polymerase by CPP32 and by S/M extracts. We also found that N-(acetyltyrosinylvalinyl-N epsilon-biotinyllysyl)aspartic acid [(2,6-dimethylbenzoyl)oxy]methyl ketone, which derivatizes the larger subunit of active ICE, can affinity label up to five active IRPs in S/M extracts. Together, these observations indicate that the processing of nuclear proteins in apoptosis involves multiple IRPs having distinct preferences for their apoptosis-associated substrates.