Comparative Purification and Characterization of Two HIN Domains, Hematopoietic Interferon-Inducible Nuclear Antigens with a 200-Amino-Acid Repeat, in Murine AIM2-Like Receptors
Comparative Purification and Characterization of Two HIN Domains, Hematopoietic Interferon-Inducible Nuclear Antigens with a 200-Amino-Acid Repeat, in Murine AIM2-Like Receptors
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小鼠 AIM2 样受体中两个 HIN 结构域（具有 200 个氨基酸重复序列的造血干扰素诱导核抗原）的比较纯化和表征
DOI：
10.1271/bbb.130544
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发表时间：
2013-11-01
影响因子：
1.6
通讯作者：
Wu, Jia-Wei
中科院分区：
文献类型：
--
作者：
Li, He;Wang, Zhi-Xin;Wu, Jia-Wei
HIN（具有 200 个氨基酸重复的造血干扰素诱导核抗原）结构域存在于所有 AIM2 样受体 (ALR) 中，并介导蛋白质/DNA 相互作用以调节先天免疫。我们使用大肠杆菌 JM109 (DE3) 宿主细胞克隆、表达和纯化人干扰素诱导蛋白 p202 (Ifi202) HINb 结构域和鼠 Ifi203 HIN 结构域。 Ifi203 HIN 结构域是溶液中的单体，具有结合双链 DNA 和 RNA 的能力。相比之下，Ifi202 HINb 结构域在溶液中是二聚体，未检测到与双链 DNA 或 RNA 的相互作用。制备Ifi203 HIN结构域和双链DNA的复合物，并获得该复合物的晶体。为了进一步分析 Ifi202 HINb 结构域的二聚体界面和 Ifi203 HIN 结构域的 DNA 结合位点，开发了两种蛋白质的模型。这项工作为理解 HIN 结构域的结构和功能提供了基础。
HIN (hematopoietic interferon-inducible nuclear antigens with a 200-amino-acid repeat) domains are found in all AIM2-like receptors (ALRs) and mediate protein/DNA interactions to regulate innate immunity. We cloned, expressed, and purified the human interferon-inducible protein p202 (Ifi202) HINb domain and the murine Ifi203 HIN domain using Escherichia coli JM109 (DE3) host cells. The Ifi203 HIN domain is a monomer in solution, and it has the ability to bind both double-stranded DNA and RNA. In contrast, the Ifi202 HINb domain is a dimer in solution, and no interaction with double-stranded DNA or RNA was detected. A complex of the Ifi203 HIN domain and double-stranded DNA was prepared, and crystals of the complex were obtained. To analyze further the dimeric interface of the Ifi202 HINb domain and the DNA binding site of the Ifi203 HIN domain, models of both proteins were developed. This work provides a basis for understanding the structure and function of HIN domains.