| Biochemical Analysis of Caspase-8-dependent Proteolysis of IRF3 in Virus-infected Cells
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Author:
Date:
2016-11-20
[Abstract] Interferon regulatory factor 3 (IRF3) is a transcription factor, which is critical for the antiviral response against a wide range of viruses (Hiscott, 2007; Ikushima et al., 2013). It gets activated in virus-infected cells via Toll like receptors (TLRs), RIG-I (retinoic acid inducible gene 1) like receptors (RLRs), cyclic GMP-AMP synthase (cGAS) – stimulator of interferon genes (STING), which are sensors of viral components in the cells (Chattopadhyay and Sen, 2014a; 2014b; Hiscott, 2007). IRF3 is a cytoplasmic protein, upon activation by virally activated sensors it gets phosphorylated, translocated to the nucleus and binds to the interferon-sensitive response element (ISRE) of the gene promoters to induce their transcription (Hiscott, 2007). IRF3 has other functions, including ...
[摘要] 干扰素调节因子3（IRF3）是一种转录因子，对于广泛病毒的抗病毒反应至关重要（Hiscott，2007; Ikushima等，2013）。通过Toll样受体（TLR），RIG-I（视黄酸诱导型基因1）受体（RLR），环状GMP-AMP合成酶（cGAS） - 干扰素基因刺激剂（STING），其在病毒感染的细胞中被激活，其中是细胞中病毒组分的传感器（Chattopadhyay和Sen，2014a; 2014b; Hiscott，2007）。 IRF3是一种细胞质蛋白，当被病毒激活的传感器激活时，其被磷酸化，转移到细胞核并与基因启动子的干扰素敏感反应元件（ISRE）结合以诱导其转录（Hiscott，2007）。 IRF3具有其他功能，包括直接刺激病毒感染细胞凋亡。在该途径中，不需要IRF3的转录活性（Chattopadhyay等，2013b; Chattopadhyay等，2016; Chattopadhyay等，2010; Chattopadhyay和Sen，2010; Chattopadhyay等，2011）。这些途径受宿主因素以及病毒的负调控。我们的研究表明IRF3可以通过caspase-8依赖性切割进行蛋白水解加工（Sears等，2011）。 ...
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| Protein Immunoprecipitation Using Nicotiana benthamiana Transient Expression System
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Author:
Date:
2015-07-05
[Abstract] Nicotiana benthamiana (N. benthamiana) is a useful model system to transiently express protein at high level. This protocol describes in detail how to transiently express protein in N. benthamiana and how to carry out protein immunoprecipitation in this expression system. This protocol can be broadly used for investigation on protein-protein interaction, protein purification and other related protein assay.
[摘要] 本塞姆氏烟草 （ ）是一种用于高水平瞬时表达蛋白质的有用的模型系统。 该协议详细描述了如何在N中瞬时表达蛋白质。 本bent和如何在此表达系统中进行蛋白质免疫沉淀。 这个协议可以广泛用于调查蛋白质 - 蛋白质相互作用，蛋白质纯化和其他相关的蛋白质测定。
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| In vitro Lipid Transfer Assay
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Author:
Date:
2013-05-05
[Abstract] This is a protocol to detect lipid transfer activity of NRF-5, a member of the LPS binding/lipid transfer protein family. The lipid transfer activity is examined by using isotope-labeled cholesterol and liposomes, and tested in two directions (Figure 1): from proteins to liposomes and from liposomes to proteins.
[摘要] 这是一种检测NRF-5（LPS结合/脂质转移蛋白家族成员）的脂质转移活性的方案。 通过使用同位素标记的胆固醇和脂质体检查脂质转移活性，并在两个方向（图1）测试：从蛋白质到脂质体，从脂质体到蛋白质。
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