由干扰素基因刺激蛋白（STING）检测环状二核苷酸
Detection of Cyclic Dinucleotides by STING.
作者信息
Du Xiao-Xia, Su Xiao-Dong
机构信息
Biodynamic Optical Imaging Center (BIOPIC), School of Life Sciences, Peking University, Yiheyuan Road No. 5, Haidian District, Beijing, 100871, China.
State Key Laboratory of Protein and Plant Gene Research, School of Life Sciences, Peking University, Yiheyuan Road No. 5, Haidian District, Beijing, 100871, China.
出版信息
Methods Mol Biol. 2017;1657:59-69. doi: 10.1007/978-1-4939-7240-1_6.
STING (stimulator of interferon genes) is an essential signaling adaptor protein mediating cytosolic DNA-induced innate immunity for both microbial invasion and self-DNA leakage. STING is also a direct receptor for cytosolic cyclic dinucleotides (CDNs), including the microbial secondary messengers c-di-GMP (3',3'-cyclic di-GMP), 3',3'cGAMP (3',3'-cyclic GMP-AMP), and mammalian endogenous 2',3'cGAMP (2',3'-cyclic GMP-AMP) synthesized by cGAS (cyclic GMP-AMP synthase). Upon CDN binding, STING undergoes a conformational change to enable signal transduction by phosphorylation and finally to active IRF3 (Interferon regulatory factor 3) for type I interferon production. Here, we describe some experimental procedures such as Isothermal Titration Calorimetry and luciferase reporter assays to study the CDNs binding and activity by STING proteins.
干扰素基因刺激蛋白（STING）是一种重要的信号衔接蛋白，介导胞质DNA诱导的针对微生物入侵和自身DNA泄漏的固有免疫。STING也是胞质环二核苷酸（CDN）的直接受体，包括微生物第二信使环二鸟苷酸（c-di-GMP，3',3'-环二鸟苷酸）、3',3'-环鸟苷酸腺苷酸（3',3'-cGAMP）以及由环磷酸鸟苷-腺苷酸合成酶（cGAS）合成的哺乳动物内源性2',3'-环鸟苷酸腺苷酸（2',3'-cGAMP）。在与CDN结合后，STING发生构象变化，通过磷酸化实现信号转导，最终激活干扰素调节因子3（IRF3）以产生I型干扰素。在此，我们描述了一些实验方法，如等温滴定量热法和荧光素酶报告基因检测，以研究STING蛋白与CDN的结合及活性。
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