Spider acetylcholine binding proteins: An alternative model to study the interaction between insect nAChRs and neonicotinoids
Spider acetylcholine binding proteins: An alternative model to study the interaction between insect nAChRs and neonicotinoids
复制标题
蜘蛛乙酰胆碱结合蛋白：研究昆虫 nAChR 和新烟碱类杀虫剂之间相互作用的替代模型
DOI：
10.1016/j.ibmb.2017.09.014
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发表时间：
2017
影响因子：
3.8
通讯作者：
Liu Zewen
中科院分区：
文献类型：
--
作者：
Bao Haibo;Meng Xiangkun;Liu Zewen
乙酰胆碱结合蛋白（Acetylcholine binding protein, achbp）是烟碱乙酰胆碱受体（nictinic Acetylcholine receptor, nAChRs）胞外结构域的同源物，是研究烟碱乙酰胆碱受体（nAChRs）的模型。特别是，由于难以异源表达而受到限制的无脊椎动物nachr的研究受益于achbp的发现。迄今为止，仅在水生软体动物中发现了achbp，而水生软体动物对新烟碱类杀虫剂（针对昆虫nAChRs的杀虫剂）的敏感性较低。然而，基于序列和组织表达分析，在蜘蛛中也发现了achbp。本文报道了水稻害虫天敌pardosapseudoannulata中的5个AChBP亚基。与软体动物的AChBP亚基相比，蜘蛛与昆虫和哺乳动物的AChBP亚基具有更高的序列相似性。p的AChBP1亚基。假环蛋白（Pp-AChBP）在Sf9细胞中表达。还表达了来自停滞林的Ls-AChBP以进行比较。在这两种achbp中，每个亚基的一个配体位点存在于两个相邻亚基之间的每个界面上。新烟碱类药物对Pp-AChBP的亲和度（kdorkivalues为7.9 ~ 18.4倍）高于对Ls-AChBP的亲和度，而依比替丁和α-班加罗毒素对Ls-AChBP的亲和度更高。这些结果表明，蜘蛛AChBP可以作为研究昆虫nAChRs与新烟碱相互作用的替代模型。
Acetylcholine binding proteins (AChBPs) are homologs of extracellular domains of nicotinic acetylcholine receptors (nAChRs) and serve as models for studies on nAChRs. Particularly, studies on invertebrate nAChRs that are limited due to difficulties in their heterologous expression have benefitted from the discovery of AChBPs. Thus far, AChBPs have been characterized only in aquatic mollusks, which have shown low sensitivity to neonicotinoids, the insecticides targeting insect nAChRs. However, AChBPs were also found in spiders based on the sequence and tissue expression analysis. Here, we report five AChBP subunits inPardosapseudoannulata, a predator enemy against rice insect pests. Spider AChBP subunits shared higher sequence similarities with nAChR subunits of both insects and mammals compared with mollusk AChBP subunits. The AChBP1 subunit ofP. pseudoannulata(Pp-AChBP) was then expressed in Sf9 cells. The Ls-AChBP fromLymnaea stagnaliswas also expressed for comparison. In both AChBPs, one ligand site per subunit was present at each interface between two adjacent subunits. Neonicotinoids had higher affinities (7.9–18.4 times based onKdorKivalues) for Pp-AChBP than for Ls-AChBP, although epibatidine and α-bungarotoxin showed higher affinities for Ls-AChBP. These results indicate that spider AChBP could be used as an alternative model to study the interaction between insect nAChRs and neonicotinoids.