烟草粉螟性信息素结合蛋白EeluPBP的表达及结合特性
Expression and binding properties of pheromone binding protein EeluPBP in Ephestia elutella
-
摘要:
为明确烟草粉螟对性信息素的识别机制，在大肠杆菌中表达了烟草粉螟性信息素结合蛋白EeluPBP，通过荧光竞争结合试验分析EeluPBP蛋白与烟草粉螟2种性信息素组分（顺，反）-9, 12-十四碳二烯-1-醇和（顺，反）-9, 12-十四碳二烯-1-醇乙酸酯的结合能力。结果表明：①EeluPBP蛋白在大肠杆菌中成功表达，蛋白质分子量大小约为15 kDa。②EeluPBP蛋白与荧光探针1-NPN的结合解离常数Kd值为0.83 μmol/L，表明体外表达的EeluPBP蛋白具有较好的活性。③EeluPBP蛋白与烟草粉螟性信息素组分（顺，反）-9, 12-十四碳二烯-1-醇具有较强的结合能力（竞争解离常数Ki值为3.26 μmol/L），表明性信息素结合蛋白EeluPBP在烟草粉螟感受性信息素的过程中发挥重要作用。
Abstract:To understand the sex pheromone perception mechanisms in Ephestia elutella, a pheromone binding protein (EeluPBP) from Ephestia elutellawas expressed in Escherichia coli in vitro. The binding capacity of EeluPBP protein with the two sex pheromones of E. elutella, (Z,E)-9, 12-tetradecadien-1-ol and (Z,E)-9, 12-tetradecadien-1-ol acetate, was evaluated by a fluorescence competitive binding assay. The results showed that: 1) EeluPBP protein was successfully expressed in E. coliwith a molecular weight of approximately 15 kDa. 2) The binding dissociation constant Kd value of EeluPBP protein with fluorescent probe 1-NPN was 0.83 μmol/L, indicating that the EeluPBP protein expressed and purified in vitro had good binding activities. 3) The recombinant EeluPBP protein had a strong binding capacity with (Z,E)-9, 12-tetradecadien-1-ol with a Ki value of 3.26 μmol/L, indicating that the pheromone binding protein EeluPBP played an important role in the perception of sex pheromone in E. elutella.
下载: