植物免疫信号分子NHP结合蛋白的虚拟筛选  

Virtual Screening of Plant Immune Signaling Molecule NHP Binding Proteins

作　　者：黄财 刘诗敏 何聪慧 蒋锌泉 苏子惠 杨婧艺 傅琼 明振华 HUANG Cai;LIU Shimin;HE Conghui;JIANG Xinquan;SU Zihui;YANG Jingyi;FU Qiong;MING Zhenhua(College of Life Science and Technology,State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources,Guangxi Key Laboratory for Sugarcane Biology,Guangxi University,Nanning,530004)

机构地区：[1]广西大学生命科学与技术学院,亚热带农业生物资源保护与利用国家重点实验室,广西甘燕生物学重点实验室,南宁530004

出　　处：《基因组学与应用生物学》2025年第2期134-143,共10页Genomics and Applied Biology

基　　金：广西自然科学基金项目(2020GXNSFFA297007);广西研究生教育创新计划资助项目(YCBZ2024012);国家级大学生创新创业训练计划项目(202310593122)共同资助。

摘　　要：植物代谢产物N-羟基哌啶酸(N-hydroxy-pipecolic acid,NHP)是植物系统获得性抗性(systemic acquired resistance,SAR)中关键的移动信号分子。然而,NHP的受体蛋白及调控其代谢、修饰和运输的关键蛋白仍不明确。本研究针对拟南芥(Arabidopsis thaliana)中受NHP诱导高表达的1883个基因,通过AlphaFill数据库筛选具有小分子结合口袋的候选蛋白;运用Chimera 1.17.3软件和DOCK 6.9软件构建结合口袋库;并通过DOCK 6.9软件筛选NHP结合能高于四组对照复合体(FMO1-NHP、UGT76B1-NHP、FMO1-Pip、UGT76B1-Pip)的结合口袋;使用Metascape数据库对预测结合蛋白进行GO功能富集及KEGG通路分析。本研究共筛选出59个可与NHP结合的蛋白口袋,对应52种蛋白质。其中,NHP与细胞色素P450类蛋白、免疫信号转导受体激酶以及关键的免疫响应蛋白激酶具有较强的结合能力。GO富集分析显示,这些蛋白主要参与了病原体刺激的防御响应、热刺激响应、蛋白质折叠和成熟调控等生物过程;分子功能方面则涉及小分子结合、水解酶活性及糖基转移酶活性等;细胞组分则富集于分泌小泡、内质网蛋白复合物及转录中介体复合物等。KEGG通路分析主要涉及内质网中蛋白质加工途径信号通路。综上所述,本研究构建了相对精准的NHP结合口袋库,CYP721A1、CRK2、MPK11和AIG1/IAN8等蛋白可能参与NHP的修饰以及SAR引发的植物免疫调控。The plant metabolite N-hydroxy-pipecolic acid(NHP)is a crucial mobile signaling molecule in systemic acquired resistance(SAR)in plants.However,the receptor for NHP,as well as the key proteins involved in its metabolism,modification,and transport,remain unclear.To address this,proteins containing small-molecule binding pockets,which are highly expressed upon NHP induction in Arabidopsis thaliana,were retrieved from the AlphaFill database.Utilizing Chimera 1.17.3 and DOCK 6.9,we created a library of binding pockets and screened for potential NHP-binding sites with higher binding energies compared to four control complexes(FMO1-NHP,UGT76B1-NHP,FMO1-Pip,UGT76B1-Pip).Gene Ontology(GO)function enrichment and KEGG pathway analyses were conducted on the predicted binding proteins via the Metascape database.In total,59 NHP-binding pockets corresponding to 52 proteins were identified.Notably,NHP exhibited the highest binding affinity for cytochrome P450 proteins,immune signaling receptor kinases,and key immune response protein kinases.GO enrichment analyses indicated that these proteins are involved in biological processes such as pathogen defense responses,heat stimulus responses,and regulation of protein folding and maturation.The molecular functions of these proteins include small-molecule binding,hydrolase activity,and glycosyltransferase activity.With cellular components identified as secretory vesicles,endoplasmic reticulum protein complexes,and transcription mediator complexes.KEGG pathway analysis primarily highlights protein processing pathways in the endoplasmic reticulum.In summary,this study constructed a comprehensive NHP binding pocket library and suggested that CYP721A1,CRK2,MPK11,and AIG1/IAN8 may participate in NHP modification and the regulation of plant immune responses during SAR.

关 键 词：系统获得性抗性(SAR) N-羟基哌啶酸(NHP) 植物免疫 分子对接 

分 类 号：S432.1[农业科学—植物病理学]