Hu Xiaohua, Huang Xianhui, Liu Jiannan, Zhao Guohui, Wu Songqing, Yu Xiaoqiang, Xu Lei, Guan Xiong, Zhang Lingling
College of Oceanology and Food Science, Quanzhou Normal University, Quanzhou 362000, China.
State Key Laboratory of Ecological Pest Control for Fujian and Taiwan Crops, School of Life Science, Fujian Agriculture and Forestry University, Fuzhou 350002, China.
Toxins (Basel). 2025 Sep 6;17(9):451. doi: 10.3390/toxins17090451.
a crucial vector mosquito that transmits many diseases that cause millions of deaths worldwide, can be controlled with subsp. (Bti). The larvicidal activity of Bti against is due primarily to Cry4Aa, Cry4Ba, and Cry11Aa, and Cyt1Aa, a protein that synergizes the activity of the Cry proteins. Interestingly, Galectins-6 and Galectins-14, members of a family of β-galactoside-binding proteins that play a role in immune responses insects, have been shown to decrease the activity of Bti toxins. The activity of other Galectins, particularly Galectin-8A, against the Cry proteins is not known. Toward this end, we cloned the gene coding for galactin-8A and expressed the recombinant protein and purified protein. The bioassay results indicated that Galectin-8A can also reduce the toxicity of Cry11Aa, but it was much stronger than Galectin-6. To investigate the interactions among Galectin-8A, Cry11Aa, and toxin receptors, Octet Red System analysis, Western blot, far-Western blot, and ELISA assay were also performed. The Octet Red System result showed that Galectin-8A could also bind to BBMVs of , with a lower kDa value than that of Galectin-6, indicating that Galectin-8A had a stronger binding affinity to BBMVs than Galectin-6. Western blot, far-Western blot, and ELISA assay analyses also demonstrated that Galectin-8A bound to receptor ALP1 and APN2, consistent with the protein docking simulation results. These findings support the conclusion that Galectin-8A blocks with ALP1 and APN2 more effectively than Galectin-6, which may subsequently reduce the toxicity of Cry11Aa in .
一种传播多种疾病并在全球导致数百万人死亡的关键病媒蚊子，可以用以色列亚种（Bti）进行控制。Bti对该蚊子的杀幼虫活性主要归因于Cry4Aa、Cry4Ba和Cry11Aa，以及Cyt1Aa，一种能增强Cry蛋白活性的蛋白质。有趣的是，半乳糖凝集素-6和半乳糖凝集素-14是β-半乳糖苷结合蛋白家族的成员，在昆虫免疫反应中发挥作用，已被证明会降低Bti毒素的活性。其他半乳糖凝集素，特别是半乳糖凝集素-8A，对Cry蛋白的活性尚不清楚。为此，我们克隆了编码半乳糖凝集素-8A的基因，并表达了重组蛋白和纯化蛋白。生物测定结果表明，半乳糖凝集素-8A也能降低Cry11Aa的毒性，但比半乳糖凝集素-6强得多。为了研究半乳糖凝集素-8A、Cry11Aa和毒素受体之间的相互作用，还进行了Octet Red系统分析、蛋白质印迹、远蛋白质印迹和酶联免疫吸附测定。Octet Red系统结果表明，半乳糖凝集素-8A也能与该蚊子的刷状缘膜囊泡（BBMVs）结合，其千道尔顿值低于半乳糖凝集素-6，表明半乳糖凝集素-8A对BBMVs的结合亲和力比半乳糖凝集素-6更强。蛋白质印迹、远蛋白质印迹和酶联免疫吸附测定分析也表明，半乳糖凝集素-8A与该蚊子的受体碱性磷酸酶1（ALP1）和氨肽酶N2（APN2）结合，这与蛋白质对接模拟结果一致。这些发现支持以下结论：半乳糖凝集素-8A比半乳糖凝集素-6更有效地与ALP1和APN2结合，这可能随后降低Cry11Aa在该蚊子中的毒性。
