Chanu Ng Kunjarani, Mandal Madan Kumar, Singh Shilpi, Yadav Mukesh Kumar, Singh Prashant Kumar, Chaurasia Neha
Environmental Biotechnology Laboratory, Department of Biotechnology and Bioinformatics, North-Eastern Hill University, Meghalaya, 793022, Shillong, India.
Department of Botany, Centre of Advanced Study in Botany, Institute of Science, Banaras Hindu University, Varanasi, 221005, India.
Arch Microbiol. 2025 Oct 24;207(12):328. doi: 10.1007/s00203-025-04491-0.
The present study focuses on the response of microalgae Graesiella emersonii NC-M1 to alpha-cypermethrin exposure at the molecular level using 2-D gel electrophoresis coupled with mass spectrometry and in-silico analysis. The proteins such as NAD(P)H-quinone oxidoreductase subunit I (+ 1.61), Heat shock proteins 70 (+ 3.01), Cytochrome P450 (+ 2.81), ABC transporters (+ 1.68), Benzoate carboxyl methyltransferase (+ 5.13), and 6(G)-Fructosyltransferase (- 3.95) were advocated as key players against alpha-cypermethrin. The accumulation pattern of these proteins was validated by a gene-expression study using qPCR. Furthermore, in-silico analysis was constructed using 3D modelling of the selected proteins, followed by the model quality assessment. The binding energy between most selected proteins and alpha-cypermethrin shows a strong interaction, except for protein benzoate carboxyl methyltransferase. Docking the ligand and the receptor offers strong binding energy, but the selected ligand, alpha-cypermethrin, is binding at the surface groove/surface-exposed binding pocket or allosteric site of the protein that can modulate other functions. The binding of alpha-cypermethrin on the selected proteins might trigger some defensive mechanism, causing them to up-accumulate under stress. Further, qPCR and docking studies supported the down-accumulation of 6(G)-fucosyltransferase (6G-FT). Taken together, these proteins were involved in the detoxification of the insecticide, DNA damage repair, and maintaining cellular homeostasis, thereby liberating the G. emersonii NC-M1 from stress conditions.
本研究聚焦于微藻艾默生格氏藻NC-M1在分子水平上对氯氰菊酯暴露的响应，采用二维凝胶电泳结合质谱分析和计算机模拟分析。诸如NAD(P)H-醌氧化还原酶亚基I（+1.61）、热休克蛋白70（+3.01）、细胞色素P450（+2.81）、ABC转运蛋白（+1.68）、苯甲酸羧基甲基转移酶（+5.13）和6(G)-果糖基转移酶（-3.95）等蛋白质被认为是对抗氯氰菊酯的关键因子。通过使用qPCR的基因表达研究验证了这些蛋白质的积累模式。此外，利用所选蛋白质的三维建模构建计算机模拟分析，随后进行模型质量评估。除了蛋白质苯甲酸羧基甲基转移酶外，大多数所选蛋白质与氯氰菊酯之间的结合能显示出强烈的相互作用。对接配体和受体提供了强大的结合能，但所选配体氯氰菊酯在蛋白质的表面凹槽/表面暴露的结合口袋或变构位点结合，可调节其他功能。氯氰菊酯与所选蛋白质的结合可能触发一些防御机制，导致它们在应激下积累增加。此外，qPCR和对接研究支持6(G)-岩藻糖基转移酶（6G-FT）的积累减少。综上所述，这些蛋白质参与了杀虫剂的解毒、DNA损伤修复和维持细胞内稳态，从而使艾默生格氏藻NC-M1从应激条件中解脱出来。
