The cleaved-Caspase-3 antibody is a marker of Caspase-9-like DRONC activity in Drosophila.
The cleaved-Caspase-3 antibody is a marker of Caspase-9-like DRONC activity in Drosophila.
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DOI：
10.1038/cdd.2009.185
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发表时间：
2010-03
影响因子：
12.4
通讯作者：
Bergmann, A.
中科院分区：
文献类型：
--
作者：
Fan, Y.;Bergmann, A.
切割的Caspase-3抗体是果蝇细胞凋亡研究中的常用工具。由于抗体是针对裂解的人胱天蛋白酶-3产生的，因此假定其检测果蝇中裂解的DRICE和DCP-1、胱天蛋白酶-3样效应物胱天蛋白酶。然而，如这里所示，强免疫反应性持续存在于drICE和dcp-1的双重突变的凋亡模型中。相反，溶酶体组分DRONC（半胱天冬酶-9样）和方舟（Apaf-1相关）的突变体不标记有切割的半胱天冬酶-3抗体。通过肽阻断实验和进一步的遗传学研究，我们提供了证据表明，切割的Caspase-3抗体识别多种蛋白质，包括DCP-1和可能的DRICE，但也至少有一个额外的未知蛋白质，所有这些都需要DRONC表位暴露。未知底物可能参与DRONC的非凋亡功能。因为切割的半胱天冬酶-3抗体不仅检测果蝇中的切割的半胱天冬酶-3样蛋白，而且还以DRONC依赖性方式检测其他蛋白，所以将切割的半胱天冬酶-3抗体视为DRONC活性的标志物而不是效应物半胱天冬酶活性更准确。
The cleaved-Caspase-3 antibody is a popular tool in apoptosis research in Drosophila. Because the antibody was raised against cleaved human Caspase-3, it was assumed that it detects cleaved DRICE and DCP-1, Caspase-3-like effector caspases in Drosophila. However, as shown here strong immunoreactivity persists in apoptotic models doubly mutant for drICE and dcp-1. In contrast, mutants of the apoptosome components DRONC (Caspase-9-like) and ARK (Apaf-1 related) do not label with cleaved-Caspase-3 antibody. By peptide blocking experiments and further genetic studies, we provide evidence that the cleaved-Caspase-3 antibody recognizes multiple proteins including DCP-1 and likely DRICE, but also at least one additional unknown protein, all of which require DRONC for epitope exposure. The unknown substrate may be involved in non-apoptotic functions of DRONC. Because the cleaved-Caspase-3 antibody not only detects cleaved Caspase-3-like proteins in Drosophila, but also other proteins in a DRONC-dependent manner, it is more accurate to consider the cleaved-Caspase-3 antibody as a marker for DRONC activity, rather than effector caspase activity.
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影响因子：
12.4
作者：
Srivastava, M.;Scherr, H.;Bergmann, A.
通讯作者：
Bergmann, A.
影响因子：
11.8
作者：
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10.5
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21.3
作者：
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作者：
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