Zhang Fangmei, Merchant Austin, Zhao Zhibin, Zhang Yunhui, Zhang Jing, Zhang Qingwen, Wang Qinghua, Zhou Xuguo, Li Xiangrui
Henan Provincial South Henan Crop Pest Green Prevention and Control Academician Workstation, Xinyang Agriculture and Forestry University, Xinyang, China.
State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing, China.
Front Physiol. 2020 Apr 1;11:212. doi: 10.3389/fphys.2020.00212. eCollection 2020.
Insect Odorant-Binding Proteins (OBPs) play crucial roles in the discrimination, binding and transportation of odorants. Herein, the full-length cDNA sequence of Minus-C OBP1 (MaltOBP1) from the Japanese pine sawyer beetle, , was cloned by 3' and 5' RACE-PCR and analyzed. The results showed that MaltOBP1 contains a 435 bp open reading frame (ORF) that encodes 144 amino acids, including a 21-amino acid signal peptide at the N-terminus. The matured MaltOBP1 protein possesses a predicted molecular weight of about 14 kDa and consists of six α-helices, creating an open binding pocket, and two disulfide bridges. Immunoblotting results showed that MaltOBP1 was most highly expressed in antennae in both sexes, followed by wings and legs. Fluorescence assays demonstrated that MaltOBP1 protein exhibited high binding affinity with (R)-(+)-α-pinene, (-)-β-pinene, -caryophyllene, (R)-(+)-limonene and (-)-verbenone, which are the main volatile compounds of the pine tree. Our combined results suggest that MaltOBP1 plays a role in host seeking behavior in .
昆虫气味结合蛋白（OBPs）在气味分子的识别、结合和运输过程中发挥着关键作用。在此，通过3'和5' RACE-PCR克隆并分析了来自日本松材线虫的Minus-C OBP1（MaltOBP1）的全长cDNA序列。结果表明，MaltOBP1包含一个435 bp的开放阅读框（ORF），编码144个氨基酸，在N端包括一个21个氨基酸的信号肽。成熟的MaltOBP1蛋白预测分子量约为14 kDa，由六个α螺旋组成，形成一个开放的结合口袋，以及两个二硫键。免疫印迹结果表明，MaltOBP1在两性的触角中表达最高，其次是翅膀和腿部。荧光分析表明，MaltOBP1蛋白与松树的主要挥发性化合物（R）-（+）-α-蒎烯、（-）-β-蒎烯、β-石竹烯、（R）-（+）-柠檬烯和（-）-马鞭草烯酮表现出高结合亲和力。我们的综合结果表明，MaltOBP1在日本松材线虫的寄主寻找行为中发挥作用。
