CED-4 forms a 2:2 heterotetrameric complex with CED-9 until specifically displaced by EGL-1 or CED-13
CED-4 forms a 2:2 heterotetrameric complex with CED-9 until specifically displaced by EGL-1 or CED-13
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DOI：
10.1038/sj.cdd.4401762
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发表时间：
2006-03-01
影响因子：
12.4
通讯作者：
Colman, PM
中科院分区：
文献类型：
--
作者：
Fairlie, WD;Perugini, MA;Colman, PM
秀丽隐杆线虫中细胞死亡的途径已经确立。在经历凋亡的细胞中，Bcl-2同源结构域3（BH 3）-唯一蛋白EGL-1在线粒体膜处结合CED-9以引起CED-4的释放，CED-4寡聚化并促进半胱天冬酶CED-3的活化。然而，尽管有许多研究，CED-4/CED-9复合物的生物物理特征尚未完全表征。在这里，我们报告的纯化的可溶性和稳定的2：2异源四聚体复合物形成的重组CED-4和CED-9共表达在细菌中。与先前的研究一致，对应于蠕虫仅BH 3蛋白（EGL-1，CED-13）的BH 3结构域的合成肽使CED-4与CED-9解离，但不与功能获得性CED-9（G169 E）突变体解离。令人惊讶的是，蠕虫BH 3结构域解离CED-4的能力是特异性的，因为哺乳动物BH 3-only蛋白不能这样做。
The pathway to cell death in Caenorhabditis elegans is well established. In cells undergoing apoptosis, the Bcl-2 homology domain 3 (BH3)-only protein EGL-1 binds to CED-9 at the mitochondrial membrane to cause the release of CED-4, which oligomerises and facilitates the activation of the caspase CED-3. However, despite many studies, the biophysical features of the CED-4/CED-9 complex have not been fully characterised. Here, we report the purification of a soluble and stable 2 : 2 heterotetrameric complex formed by recombinant CED-4 and CED-9 coexpressed in bacteria. Consistent with previous studies, synthetic peptides corresponding to the BH3 domains of worm BH3-only proteins (EGL-1, CED-13) dissociate CED-4 from CED-9, but not from the gain-of-function CED-9 (G169E) mutant. Surprisingly, the ability of worm BH3 domains to dissociate CED-4 was specific since mammalian BH3-only proteins could not do so.