Rapid inactivation of a moth pheromone
Rapid inactivation of a moth pheromone
复制标题
DOI：
10.1073/pnas.0505340102
复制
发表时间：
2005-09-27
影响因子：
11.1
通讯作者：
Leal, WS
中科院分区：
文献类型：
--
作者：
Ishida, Y;Leal, WS
本研究从野蚕（Antheraea polyphemus PDE（ApolPDE））中分离、克隆并表达了一种雄蛾触角特异性信息素降解酶（PDE）。polyphemus，这似乎是必不可少的快速失活的信息素在飞行过程中。酶活性在蛹期的第13天开始检测，在成虫期的第2天达到峰值。通过多个色谱步骤从第2天雄性触角分离的ApolPDE的从头测序导致cDNA克隆。纯化的重组ApolPDE，杆状病毒表达，迁移与天然蛋白质的天然聚丙烯酰胺和等电聚焦凝胶电泳相同的流动性。感觉淋巴中ApolPDE（0.5 μ M）的浓度比信息素结合蛋白的浓度低约20，000 μ M。天然和重组ApolPDE表现出相当的动力学参数，营业额类似的羧肽酶和底物特异性略低于乙酰胆碱酯酶。信息素的快速失活，甚至比以前估计的更快，在动力学上与持续的气味介导的飞蛾飞行所需的时间分辨率兼容。
We have isolated, cloned, and expressed a male antennae-specific pheromone-degrading enzyme (PDE) [Antheraea polyphemus PDE (ApolPDE), formerly known as Sensillar Esterase] from the wild silkmoth, A. polyphemus, which seems essential for the rapid inactivation of pheromone during flight. The onset of enzymatic activity was detected at day 13 of the pupal stage with a peak at day 2 adult stage. De novo sequencing of ApolPDE, isolated from day 2 male antennae by multiple chromatographic steps, led to cDNA cloning. Purified recombinant ApolPDE, expressed by baculovirus, migrated with the same mobility as the native protein on both native polyacrylamide and isoelectric focusing gel electrophoresis. Concentration of ApolPDE (0.5 mu M) in the sensillar lymph is approximate to 20,000 lower than that of a pheromone-binding protein. Native and recombinant ApolPDE showed comparable kinetic parameters, with turnover number similar to that of carboxypeptidase and substrate specificity slightly lower than that of acetylcholinesterase. The rapid inactivation of pheromone, even faster than previously estimated, is kinetically compatible with the temporal resolution required for sustained odorant-mediated flight in moths.