Mch3, a novel human apoptotic cysteine protease highly related to CPP32.
Mch3, a novel human apoptotic cysteine protease highly related to CPP32.
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发表时间：
1995-12
期刊：
影响因子：
11.2
通讯作者：
T. Fernandes‐Alnemri;Atsushi Takahashi;R. Armstrong;J. Krebs;L. Fritz;K. Tomaselli;Lijuan Wang;Zailin Yu;C. Croce;Guy Salveson;W. Earnshaw;G. Litwack;E. Alnemri
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作者：
T. Fernandes‐Alnemri;Atsushi Takahashi;R. Armstrong;J. Krebs;L. Fritz;K. Tomaselli;Lijuan Wang;Zailin Yu;C. Croce;Guy Salveson;W. Earnshaw;G. Litwack;E. Alnemri
最近的证据表明，与秀丽隐杆线虫 CED-3 相关的哺乳动物半胱氨酸蛋白酶是哺乳动物程序性细胞死亡或细胞凋亡的关键组成部分。我们最近表明，CPP32 和 Mch2 α 半胱氨酸蛋白酶分别裂解凋亡标记聚（ADP-核糖）聚合酶（PARP）和核纤层蛋白。在这里，我们报道了一种新的 Ced-3/白细胞介素 1 β 转换酶相关基因的克隆，命名为 Mch3，与其他家族成员相比，该基因编码与 CPP32 具有最高同源性的蛋白质。还鉴定出了一种名为 Mch3 beta 的选择性剪接异构体。细菌表达的重组 Mch3 具有内在的自催化/自激活活性。 Mch3 α 对肽底物 DEVD-7-氨基-4-甲基香豆素和 PARP 的比活性与 CPP32 相似。与白细胞介素 1 β 转换酶和 CPP32 一样，活性 Mch3 α 由源自前体 (proMch3 α) 的两个亚基组成。令人感兴趣的是，重组 CPP32-p17 亚基可以与重组 Mch3 α-p12 亚基形成活性异聚酶复合物，反之亦然，正如异聚复合物诱导 Sf9 细胞凋亡的能力所确定的。这些数据表明 proMch3 α 和 proCPP32 可以相互作用形成活性 Mch3 α/CPP32 异聚复合物。我们还提供了证据表明 CPP32 可以有效地裂解 proMch3 α，但反之则不然，这表明 Mch3 α 体内激活可能部分取决于 CPP32 活性。结构和比活性的高度保守性以及Mch3α和CPP32在同一细胞中的共存表明，细胞凋亡过程中观察到的PARP裂解活性不能仅仅归因于CPP32，也可能是Mch3α的活性。
Recent evidence suggests that mammalian cysteine proteases related to Caenorhabditis elegans CED-3 are key components of mammalian programmed cell death or apoptosis. We have shown recently that the CPP32 and Mch2 alpha cysteine proteases cleave the apoptotic markers poly(ADP-ribose) polymerase (PARP) and lamins, respectively. Here we report the cloning of a new Ced-3/interleukin 1 beta-converting enzyme-related gene, designated Mch3, that encodes a protein with the highest degree of homology to CPP32 compared to other family members. An alternatively spliced isoform, named Mch3 beta, was also identified. Bacterially expressed recombinant Mch3 has intrinsic autocatalytic/autoactivation activity. The specific activity of Mch3 alpha toward the peptide substrate DEVD-7-amino-4-methylcoumarin and PARP resembles that of CPP32. Like interleukin 1 beta-converting enzyme and CPP32, the active Mch3 alpha is made of two subunits derived from a precursor (proMch3 alpha). It was of interest that recombinant CPP32-p17 subunit can form an active heteromeric enzyme complex with recombinant Mch3 alpha-p12 subunit and vice versa, as determined by the ability of the heteromeric complexes to induce apoptosis in Sf9 cells. These data suggest that proMch3 alpha and proCPP32 can interact to form an active Mch3 alpha/CPP32 heteromeric complex. We also provide evidence that CPP32 can efficiently cleave proMch3 alpha, but not the opposite, suggesting that Mch3 alpha activation in vivo may depend in part on CPP32 activity. The high degree of conservation in structure and specific activity and the coexistence of Mch3 alpha and CPP32 in the same cell suggests that the PARP cleavage activity observed during apoptosis cannot solely be attributed to CPP32 but could also be an activity of Mch3 alpha.