Structural Analysis and Deletion Mutagenesis Define Regions of QUIVER/SLEEPLESS that Are Responsible for Interactions with Shaker-Type Potassium Channels and Nicotinic Acetylcholine Receptors.
Structural Analysis and Deletion Mutagenesis Define Regions of QUIVER/SLEEPLESS that Are Responsible for Interactions with Shaker-Type Potassium Channels and Nicotinic Acetylcholine Receptors.
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DOI：
10.1371/journal.pone.0148215
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发表时间：
2016
期刊：
影响因子：
3.7
通讯作者：
Joiner WJ
中科院分区：
文献类型：
--
作者：
Wu M;Liu CZ;Joiner WJ
Ly 6蛋白是在大多数动物中发现的内源性原毒素。它们在结构和功能上与蛇α-神经毒素有惊人的相似之处，包括离子通道和胆碱能信号的调节。然而，Ly 6蛋白调节效应分子的结构贡献知之甚少。这个问题与Ly 6蛋白质QUIVER/SLEEPLESS（QVR/SSS）特别相关，该蛋白质先前已被证明通过上调钾（K）通道和下调促醒神经元中的烟碱乙酰胆碱受体（nAChR）来抑制兴奋性和突触传递，以促进果蝇的睡眠。使用缺失诱变，共免疫沉淀，离子通量测定，表面标记和共聚焦显微镜，我们证明，只有循环2所需的许多先前描述的属性SSS转染细胞，包括与K通道和nAChRs的相互作用。总的来说，我们的数据表明，QVR/SSS，并通过扩展可能其他Ly 6蛋白，目标效应分子使用有限的蛋白基序。映射这些图案可能是有用的，在合理设计的药物，模仿或抑制Ly 6效应相互作用，以调节神经系统功能。
Ly6 proteins are endogenous prototoxins found in most animals. They show striking structural and functional parallels to snake α-neurotoxins, including regulation of ion channels and cholinergic signaling. However, the structural contributions of Ly6 proteins to regulation of effector molecules is poorly understood. This question is particularly relevant to the Ly6 protein QUIVER/SLEEPLESS (QVR/SSS), which has previously been shown to suppress excitability and synaptic transmission by upregulating potassium (K) channels and downregulating nicotinic acetylcholine receptors (nAChRs) in wake-promoting neurons to facilitate sleep in Drosophila. Using deletion mutagenesis, co-immunoprecipitations, ion flux assays, surface labeling and confocal microscopy, we demonstrate that only loop 2 is required for many of the previously described properties of SSS in transfected cells, including interactions with K channels and nAChRs. Collectively our data suggest that QVR/SSS, and by extension perhaps other Ly6 proteins, target effector molecules using limited protein motifs. Mapping these motifs may be useful in rational design of drugs that mimic or suppress Ly6-effector interactions to modulate nervous system function.