Detection of MLKL Oligomerization During Programmed Necrosis
Detection of MLKL Oligomerization During Programmed Necrosis
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DOI：
10.1007/978-1-4939-8754-2_8
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发表时间：
2018-01-01
期刊：
影响因子：
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通讯作者：
Liu, Zheng-Gang
中科院分区：
文献类型：
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作者：
Cai, Zhenyu;Liu, Zheng-Gang
程序性坏死，也称为坏死性凋亡，是一种由受体相互作用蛋白激酶RIP1（或RIPK1）、RIP3（或RIPK3）和混合谱系激酶结构域样蛋白MLKL介导的调节性坏死细胞死亡形式。在诱导程序性坏死后，MLKL被RIP3磷酸化并寡聚化，然后蛋白质易位到细胞质膜以执行程序性坏死。在这里，我们描述了一个详细的协议，以检测MLKL寡聚化在坏死性凋亡细胞的蛋白质印迹分析在非还原条件下。因此，我们建立了检测程序性坏死通路激活的方法。
Programmed necrosis, also known as necroptosis, is a form of regulated necrotic cell death that is mediated by receptor-interacting protein kinases RIP1 (or RIPK1), RIP3 (or RIPK3), and the mixed lineage kinase domain-like protein, MLKL. Following the induction of programmed necrosis, MLKL is phosphorylated by RIP3 and oligomerizes and then the protein translocates to cell plasma membrane in order to execute programmed necrosis. Here, we describe a detailed protocol to detect MLKL oligomerization in necroptotic cells by Western blotting analysis under nonreducing condition. Therefore, we established the method to detect the activation of programmed necrotic pathway.