Danty E, Briand L, Michard-Vanhée C, Perez V, Arnold G, Gaudemer O, Huet D, Huet J C, Ouali C, Masson C, Pernollet J C
Centre Européen des Sciences du Goût, Centre National de la Recherche Scientifique (CNRS), Unité "Olfaction, Gustation, Nutrition," 21000 Dijon, France.
J Neurosci. 1999 Sep 1;19(17):7468-75. doi: 10.1523/JNEUROSCI.19-17-07468.1999.
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.
气味结合蛋白（OBPs）是一类含量丰富的小细胞外蛋白，被认为通过携带、失活和/或选择气味刺激参与气味-信息素检测的受体外周事件。除了吸引雄蜂交配外，已知蜂王信息素在蜂群组织中起着关键作用。我们首次在一种社会性昆虫中鉴定出一种名为触角特异性蛋白1（ASP1）的结合蛋白，它能结合至少一种主要的蜂王信息素成分。通过cDNA克隆、在毕赤酵母中表达以及信息素结合对ASP1进行了表征。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。质谱分析表明重组蛋白呈现出天然蛋白的准确特征，N端测序和排阻色谱显示重组ASP1是二聚体。与另一种假定的蜜蜂OBP即ASP2相反，ASP1与蜂王信息素的主要成分相互作用。在发育过程中，通过非变性电泳跟踪ASP1的生物合成积累，其在羽化前1天开始，与嗅觉神经元的功能成熟同时发生。同工型ASP1b在工蜂中与ASP1a同时出现，但在雄蜂中仅在羽化后约2周出现。体内表达和异源表达的比较表明，ASP1同工型之间的差异可能是由于二聚化，这可能在与配偶吸引相关的生理过程中发挥作用。
