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首页> 外文期刊>The Journal of Neuroscience: The Official Journal of the Society for Neuroscience >Cloning and expression of a queen pheromone-binding protein in the honeybee: an olfactory-specific, developmentally regulated protein.
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Cloning and expression of a queen pheromone-binding protein in the honeybee: an olfactory-specific, developmentally regulated protein.

机译：蜂后信息素结合蛋白的克隆和表达：一种嗅觉特异性的发育调控蛋白。

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摘要

Odorant-binding proteins (OBPs) are small abundant extracellular proteins thought to participate in perireceptor events of odor-pheromone detection by carrying, deactivating, and/or selecting odor stimuli. The honeybee queen pheromone is known to play a crucial role in colony organization, in addition to drone sex attraction. We identified, for the first time in a social insect, a binding protein called antennal-specific protein 1 (ASP1), which binds at least one of the major queen pheromone components. ASP1 was characterized by cDNA cloning, expression in Pichia pastoris, and pheromone binding. In situ hybridization showed that it is specifically expressed in the auxiliary cell layer of the antennal olfactory sensilla. The ASP1 sequence revealed it as a divergent member of the insect OBP family. The recombinant protein presented the exact characteristics of the native protein, as shown by mass spectrometry, and N-terminal sequencing and exclusion-diffusion chromatography showed that recombinant ASP1 is dimeric. ASP1 interacts with queen pheromone major components, opposite to another putative honeybee OBP, called ASP2. ASP1 biosynthetic accumulation, followed by nondenaturing electrophoresis during development, starts at day 1 before emergence, in concomitance with the functional maturation of olfactory neurons. The isobar ASP1b isoform appears simultaneously to ASP1a in workers, but only at approximately 2 weeks after emergence in drones. Comparison of in vivo and heterologous expressions suggests that the difference between ASP1 isoforms might be because of dimerization, which might play a physiological role in relation with mate attraction.
机译：气味结合蛋白（OBP）是小的丰富的细胞外蛋白，被认为通过携带，失活和/或选择气味刺激物参与气味信息素检测的感知受体事件。众所周知，除了蜂巢性吸引外，蜜蜂女王费洛蒙在殖民地组织中也起着至关重要的作用。我们首次在社交昆虫中发现了一种结合蛋白，称为触角特异性蛋白1（ASP1），它可以结合至少一种主要的女王信息素成分。 ASP1的特征是cDNA克隆，巴斯德毕赤酵母中的表达和信息素结合。原位杂交表明它在触角嗅觉感受器的辅助细胞层中特异性表达。 ASP1序列显示它是昆虫OBP家族的一个不同成员。重组蛋白表现出天然蛋白的确切特征，如质谱分析所示，N端测序和排阻扩散色谱表明重组ASP1是二聚体。 ASP1与女王信息素主要成分相互作用，与另一种假定的蜜蜂OBP相反，称为ASP2。在嗅觉神经元功能成熟的同时，ASP1的生物合成积累以及随后在发育过程中的非变性电泳在出现前的第1天开始。在工人中，等压ASP1b同工型与ASP1a同时出现，但仅在无人机出现后约2周出现。体内和异源表达的比较表明，ASP1亚型之间的差异可能是由于二聚化，这可能与伴侣吸引有关，起着生理作用。

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