Immunoblot Analysis of Linear Polyubiquitination of NEMO.
Immunoblot Analysis of Linear Polyubiquitination of NEMO.
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NEMO 线性多泛素化的免疫印迹分析。
DOI：
10.1007/978-1-4939-2422-6_17
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发表时间：
2015
期刊：
影响因子：
--
通讯作者：
K.
中科院分区：
文献类型：
--
作者：
Sasaki;Y.;Fujita;H.;Nakai;M.;and Iwai;K.
炎性细胞因子如TNF-α和IL-1的刺激通过激活IKK复合体激活典型的NF-κB通路。IKK激活的机制已被广泛研究，泛素系统的参与已被很好地记录。我们最近报道了一种新的泛素连接酶复合物LUBAC参与IKK复合物的激活。LUBAC由一个催化亚基HOIP和两个辅助分子HOIL-1L和SHARPIN组成，并通过将线性多泛素链偶联到IKK复合物的调节亚基NEMO （IKKγ）来激活IKK复合物。在本章中，我们描述了用抗线性泛素抗体免疫印迹法检测NEMO线性多泛素化的方案。
Stimulation with inflammatory cytokines such as TNF-α and IL-1 activates the canonical NF-κB pathway through the activation of the IKK complex. The mechanism underlying IKK activation has been extensively studied and the involvement of the ubiquitin system has been well documented. We have recently reported that a novel ubiquitin ligase complex, LUBAC is involved in the activation of the IKK complex. LUBAC consists of one catalytic subunit, HOIP and two accessory molecules, HOIL-1L and SHARPIN and activates the IKK complex by conjugating the linear polyubiquitin chains to NEMO (IKKγ), the regulatory subunit of IKK complex. In this chapter, we describe the protocol for the detection of the linear polyubiquitination of NEMO by the immunoblotting using anti-linear ubiquitin antibody.