Ahmad Waqar, Rahman Ziaur, Khan Haji, Nawab Javed, Rahman Hazir, Siddiqui Muhammad Faisal, Saeed Wajeeha
Department of Microbiology, Abdul Wali Khan University Mardan, Mardan, Khyber Pakhtunkhwa, Pakistan.
Centre of Biotechnology and Microbiology, University of Swat, Swat, Khyber Pakhtunkhwa, Pakistan.
Front Plant Sci. 2024 Nov 7;15:1429890. doi: 10.3389/fpls.2024.1429890. eCollection 2024.
is a plant-pathogenic fungus and a responsible agent for causing peach leaf curl disease. affects peach fruit production and contributes to global economic losses. Commercial fungicides may provide temporary relief; however, their overuse resulted in adverse environmental consequences as well as led to drug-resistant strains of . Therefore, the discovery of novel drug targets for the future synthesis of antifungal drugs against is needed. Here we studied by computational proteomics approaches. The whole genome and proteome of . were subjected to subtractive proteomics, high-throughput virtual screening, and molecular dynamic simulations. We employed subtractive proteomics analysis of 4,659 proteins extracted from UniProtKB database; after filtering out homologous and non-essential proteins, we identified 189 essential ones, including nine that participated in the crucial metabolic pathways of the pathogen. These proteins were categorized as nuclear ( = 116), cytoplasmic ( = 37), and membrane ( = 36). Of those essential proteins, glutamate-cysteine ligase (GCL) emerged as one promising target due to its essential function for glutathione biosynthesis process which facilitates survival and pathogenicity. To validate GCL as an antifungal target, virtual screening and molecular docking studies with various commercial fungicides were carried out to better characterize GCL as a drug target. The data showed strong binding affinities for polyoxin D, fluoxastrobin, trifloxystrobin, and azoxystrobin within the active site of GCL. Polyoxin D showed a strong affinity when the measured docking score was at -7.34 kcal/mol, while molecular dynamics simulations confirmed stable interactions (three hydrogen bonds, two hydrophobic bonds, and one salt bridge interaction), supporting our findings that GCL represents an excellent target for antifungal drug development efforts. The results showed that GCL, as an innovative target for future fungicide designs to combat , provides an avenue toward creating more effective peach leaf curl disease treatments while mitigating environmental harm caused by its current use.
是一种植物病原真菌，是引起桃叶卷曲病的致病因子。它影响桃的果实产量，造成全球经济损失。商业杀菌剂可能会提供暂时的缓解；然而，它们的过度使用导致了不利的环境后果，并产生了对其具有耐药性的菌株。因此，需要发现新的药物靶点，以便未来合成针对的抗真菌药物。在这里，我们通过计算蛋白质组学方法研究了。的全基因组和蛋白质组进行了减法蛋白质组学、高通量虚拟筛选和分子动力学模拟。我们对从UniProtKB数据库中提取的4659种蛋白质进行了减法蛋白质组学分析；在筛选出同源和非必需蛋白质后，我们确定了189种必需蛋白质，其中9种参与了病原体的关键代谢途径。这些蛋白质被分类为细胞核（=116）、细胞质（=37）和膜（=36）。在这些必需蛋白质中，谷氨酸-半胱氨酸连接酶（GCL）因其在谷胱甘肽生物合成过程中的基本功能而成为一个有前景的靶点，该过程促进了的生存和致病性。为了验证GCL作为抗真菌靶点，我们与各种商业杀菌剂进行了虚拟筛选和分子对接研究，以更好地将GCL表征为药物靶点。数据显示，多氧霉素D、氟嘧菌酯、肟菌酯和嘧菌酯在GCL活性位点具有很强的结合亲和力。当测得的对接分数为-7.34 kcal/mol时，多氧霉素D显示出很强的亲和力，而分子动力学模拟证实了稳定的相互作用（三个氢键、两个疏水键和一个盐桥相互作用），支持了我们的发现，即GCL是抗真菌药物开发的一个优秀靶点。结果表明，GCL作为未来杀菌剂设计以对抗的创新靶点，为创造更有效的桃叶卷曲病治疗方法提供了一条途径，同时减轻了其当前使用所造成的环境危害。
