Expression, purification and crystallization of the N-terminal Solanaceae domain of the Sw-5b NLR immune receptor.
Expression, purification and crystallization of the N-terminal Solanaceae domain of the Sw-5b NLR immune receptor.
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DOI：
10.1107/s2053230x20016398
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发表时间：
2021-01
期刊：
影响因子：
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通讯作者：
Jia Li;Jian Xin;Xinyan Zhao;Yaqian Zhao;Tongkai Wang;Weiman Xing;Xiaorong Tao
中科院分区：
文献类型：
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作者：
Jia Li;Jian Xin;Xinyan Zhao;Yaqian Zhao;Tongkai Wang;Weiman Xing;Xiaorong Tao
植物核苷酸结合域和富亮氨酸重复序列受体在识别病原菌效应子和激活植物免疫中起着重要作用。番茄NLR Sw-5 b是一种卷曲螺旋NLR（CC-NLR）免疫受体，其赋予对番茄斑萎病毒的抗性，番茄斑萎病毒在全球范围内导致农业作物的严重经济损失。与其他CC-NLR相比，Sw-5 b具有一个扩展的茄科结构域（SD）。Sw-5 b的SD对于识别番茄斑萎病毒的病毒运动蛋白NSm至关重要。在茄科植物的许多NLR中也经常检测到SD。然而，在除茄科以外的动物或植物中未检测到与SD同源的序列。SD蛋白的性质在很大程度上是未知的，因此3D结构信息对于更好地了解其在病原体感知和免疫受体激活中的作用至关重要。本文报道了Sw-5 b SD（氨基酸1-245）的表达、纯化和结晶。SD蛋白的天然晶体和硒代蛋氨酸取代晶体均属于空间群P3112，晶胞参数分别为a = 81.53，B = 81.53，c = 98.44 nm和a = 81.63，B = 81.63，c = 98.80 nm。这是首次报道茄科植物NLR蛋白非经典SD结构域的结构研究。
Plant nucleotide-binding domain and leucine-rich repeat receptors (NLRs) play crucial roles in recognizing pathogen effectors and activating plant immunity. The tomato NLR Sw-5b is a coiled-coil NLR (CC-NLR) immune receptor that confers resistance against tospoviruses, which cause serious economic losses in agronomic crops worldwide. Compared with other CC-NLRs, Sw-5b possesses an extended N-terminal Solanaceae domain (SD). The SD of Sw-5b is critical for recognition of the tospovirus viral movement protein NSm. An SD is also frequently detected in many NLRs from Solanaceae plants. However, no sequences homologous to the SD have been detected in animals or in plants other than Solanaceae. The properties of the SD protein are largely unknown, and thus 3D structural information is vital in order to better understand its role in pathogen perception and the activation of immune receptors. Here, the expression, purification and crystallization of Sw-5b SD (amino acids 1-245) are reported. Native and selenomethionine-substituted crystals of the SD protein belonged to space group P3112, with unit-cell parameters a = 81.53, b = 81.53, c = 98.44 Å and a = 81.63, b = 81.63, c = 98.80 Å, respectively. This is the first report of a structural study of the noncanonical SD domain of the NLR proteins from Solanaceae plants.