Solution structure of NOD1 CARD and mutational analysis of its interaction with the CARD of downstream kinase RICK
Solution structure of NOD1 CARD and mutational analysis of its interaction with the CARD of downstream kinase RICK
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DOI：
10.1016/j.jmb.2006.09.067
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发表时间：
2007-01-05
影响因子：
5.6
通讯作者：
Cusack, Stephen
中科院分区：
文献类型：
--
作者：
Manon, Florence;Favier, Adrien;Cusack, Stephen
NOD1是一种胞质信号宿主模式识别受体，由caspase激活和募集结构域（CARD）、核苷酸结合和寡聚化结构域（NOD）和富含亮氨酸的重复序列组成。在识别特定的细菌配体后，它通过下游效应激酶RICK （RIP2）激活NF-kappa B通路，在先天免疫中发挥关键作用。RICK通过各自的卡牌的相互作用被NOD1招募。本文介绍了NOD1 CARD的高分辨率核磁共振结构。它通常类似于其他已知结构的卡，由六个紧密排列的螺旋组成，尽管最后一个螺旋的长度和方向是不寻常的。通过细胞裂解物的免疫沉淀和体内NF-kappa B激活，研究了NOD1和RICK CARD结构域的突变，以确定CARD-CARD相互作用和下游信号传导的重要残基。结果表明，该相互作用严重依赖于NOD1 CARD上的三个酸性残基和RICK CARD上的三个碱性残基，因此可能具有强静电成分，类似于其他特征CARD-CARD相互作用。
NOD1 is a cytosolic signalling host pattern-recognition receptor composed of a caspase-activating and recruitment domain (CARD), a nucleotide-binding and oligomerization domain (NOD) and leucine-rich repeats. It plays a crucial role in innate immunity by activating the NF-kappa B pathway via its downstream effector the kinase RICK (RIP2) following the recognition of a specific bacterial ligand. RICK is recruited by NOD1 through interaction of their respective CARDs. Here we present the high resolution NMR structure of the NOD1 CARD. It is generally similar to other CARDs of known structure, consisting of six tightly packed helices, although the length and orientation of the last helix is unusual. Mutations in both the NOD1 and RICK CARD domains were assayed by immuno-precipitation of cell lysates and in vivo NF-kappa B activation in order to define residues important for CARD-CARD interaction and downstream signalling. The results show that the interaction is critically dependent on three acidic residues on NOD1 CARD and three basic residues on RICK CARD and thus is likely to have a strong electrostatic component, similar to other characterised CARD-CARD interactions.