Identification of a new subtilisin-like protease NbSLP2 interacting with cytoskeletal protein septin in Microsporidia Nosema bombycis
Identification of a new subtilisin-like protease NbSLP2 interacting with cytoskeletal protein septin in Microsporidia Nosema bombycis
复制标题
在家蚕微孢子虫中鉴定与细胞骨架蛋白 septin 相互作用的新枯草杆菌蛋白酶样蛋白酶 NbSLP2
DOI：
10.1016/j.jip.2017.06.004
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发表时间：
2017-09-01
影响因子：
3.4
通讯作者：
Zhou, Zeyang
中科院分区：
文献类型：
--
作者：
Liu, Fangyan;Ma, Qiang;Zhou, Zeyang
家蚕微孢子虫是蚕业微粒子病的病原，每年给蚕业生产造成重大损失。类枯草杆菌蛋白酶（SLP）是丝氨酸蛋白酶的一种，与真菌的致病性有关。在这项研究中，我们确定了一个新的63.8 kDa的枯草杆菌蛋白酶样蛋白酶NbSLP 2与预测的跨膜结构域从微孢子虫，N。蚕RT-PCR结果显示，感染后第3天开始检测到NbSLP 2基因的转录。免疫荧光分析表明NbSLP 2主要分布在N.蚕免疫共沉淀和液相色谱-串联质谱（LC-MS/MS）分析表明，NbSLP 2与N. bombycis，这是一种细胞骨架蛋白。IFA显示NbSLP 2和Nbseptin 2共定位于孢子壁下。NbSLP 2可以被Nbseptin 2拉下，进一步证实了NbSLP 2和Nbseptin 2之间的相互作用。作为具有跨膜结构域的重要丝氨酸蛋白酶，NbSLP 2与Nbseptin 2相互作用，邻近膜的支架蛋白可以提供稳定NbSLP 2以用于其水解功能的优势。
Nosema bombycis is the pathogen of pebrine which brings heavy losses to sericulture every year. As a member of serine proteases, subtilisin-like protease (SLP) is related to the pathogenicity in fungi. In this study, we characterized a novel 63.8 kDa subtilisin-like protease NbSLP2 with a predicted transmembrane domain from Microsporidia, N. bombycis. RT-PCR showed that the transcript of NbSLP2 was detected from third day post infection. Immunofluorescence assay (IFA) indicated that NbSLP2 mainly scattered around the spore wall of N. bombycis. Co-immunoprecipitation data and liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) analysis revealed that NbSLP2 directly interacts with septin2 of N. bombycis, which is a cytoskeletal protein. IFA showed that NbSLP2 and Nbseptin2 co-localized beneath the spore wall. NbSLP2 can be pulled down by Nbseptin2, further confirming the interaction between NbSLP2 and Nbseptin2. As an important serine protease with a transmembrane domain, NbSLP2 interacting with Nbseptin2, a scaffold protein adjacent to the membrane may provide advantages to stabilize the NbSLP2 for its hydrolysis function.