刚地弓形虫ME49虫株ROP16蛋白的生物信息学分析  

Bioinformatics analysis of the ROP16 protein of Toxoplasma gondii strain ME49

作　　者：党甜甜 殷荷 李佳铭 田婷婷 周月 赵志军 DANG Tiantian;YIN He;LI Jiaming;TIAN Tingting;ZHOU Yue;ZHAO Zhijun(General Hospital of Ningxia Medical University,Yinchuan 750004,China;Ningxia Medical University College of Laboratory Science;Ningxia Key Laboratory of Pathogenic Microbiology)

机构地区：[1]宁夏医科大学总医院医学实验中心,宁夏银川750004 [2]宁夏医科大学检验学院 [3]宁夏临床病原微生物重点实验室

出　　处：《中国病原生物学杂志》2023年第12期1401-1406,共6页Journal of Pathogen Biology

基　　金：宁夏自然科学基金项目(No.2022AAC03547);宁夏回族自治区重点研发计划项目(2023BEG02002);宁夏医科大学总医院“新入职硕士培养”项目。

摘　　要：目的应用生物信息学方法分析弓形虫ME49虫株ROP16蛋白(ME49-ROP16)的理化性质、结构特点等,并预测其潜在的B、T细胞抗原表位及其互作蛋白,为弓形虫疫苗的研究提供理论基础。方法登录NCBI数据库下载ME49虫株ROP16蛋白的氨基酸序列,利用在线软件ProtParam预测ROP16蛋白的理化性质;使用ProScale预测ROP16蛋白的亲水性、柔韧性、表面可及性及β-折叠;采用SignalP-4.1预测ROP16蛋白的信号肽;运用TMHMM-2.0预测ROP16蛋白的跨膜结构;利用SOMPA和Phyre2预测ROP16蛋白的二、三级结构;通过Netphos3.1、NetNGlyc1.0、YinOYang-1.2预测ROP16蛋白的磷酸化位点、N-糖基化位点和O-糖基化位点;运用Predicting Antigenic Peptides预测ROP16蛋白的抗原决定簇;使用IEDB预测ROP16蛋白的优势B细胞表位;运用SYFPEITHI预测ROP16蛋白的T细胞优势表位;采用STRING预测软件分析与其相互作用的蛋白。结果ME49-ROP16蛋白由707个氨基酸组成,分子式为C_(3338)H_(5350)N_(966)O_(1033)S_(21),相对分子质量76.21631×10^(3),其理论等电点是8.97,不稳定系数为59.30,为不稳定蛋白;脂溶性指数为79.25,总的平均亲水性为-0.320,为亲水性蛋白;其二级结构中α-螺旋占35.64%,延伸链占10.75%,β-转角占5.94%,无规卷曲占47.67%;该蛋白含有83个氨基酸磷酸化位点,14个N-糖基化位点,无O-糖基化位点。ME49-ROP16蛋白具有信号肽序列,无跨膜结构;其平均抗原倾向指数为1.0340,共有25个抗原决定簇区域,有7条B细胞优势表位,14条CTL细胞优势表位,14条Th细胞优势表位,与ACC1、ACC2、PKG、ROP18为互作蛋白。结论生物信息学分析ME49-ROP16蛋白为不稳定亲水性蛋白,含有多个B、T细胞优势表位,并与ACC1、ACC2、PKG、ROP18为互作蛋白,为以该蛋白为靶点的弓形虫疫苗研制奠定了基础。Objective Bioinformatics methods were applied to analyse the physicochemical properties and structural features of the ROP16 protein of Toxoplasma gondii strain ME49(ME49-ROP16)and to predict its potential B and T cell antigenic epitopes and its intercalating proteins,so as to provide a theoretical basis for the research of Toxoplasma gondii vaccine.Methods Log in NCBI data base to search and download the amino acid sequence of ME49-ROP16.Hydrophilicity,flexibility,surface accessibilityandβ-folding of the ROP16 protein were predicted by using ProScale;SignalP-4.1 was used to predict the signal peptide of ROP16 protein;TMHMM-2.0 was used to predict the transmembrane structure of ROP16 protein;SOMPA and Phyre2 were used to predict the secondary and tertiary structure of ROP16 protein;The phosphorylation site,N-glycosylation site and O-glycosylation site of ROP16 protein were predicted by Netphos3.1,NetNGlyc1.0 and YinOYang-1.2;The antigenic determinant clusters of ROP16 protein were predicted using Predicting Antigenic Peptides;the dominant B-cell epitopes of ROP16 protein were predicted using IEDB;the dominant T-cell epitopes of ROP16 protein were predicted using SYFPEITHI;and proteins interacting with it were analysed using STRING prediction software.Results ME49-ROP16 protein consists of 707 amino acids with the molecular formula C_(3338)H_(5350)N_(966)O_(1033)S_(21) and a relative molecular mass of 76.21631×10^(3),its theoretical isoelectric point is 8.97 and its instability coefficient is 59.30,making it an unstable protein.Its lipid solubility index is 79.25,with a total average hydrophilicity of-0.320,indicating that it is a hydrophilic protein.Inthesecondarystructure,α-helix accountedfor 35.64%,extendedchainaccountedfor 10.75%,β-turnsaccountedfor 5.94%,randomcurlingaccountedfor 47.67%.The protein contains 83 amino acid phosphorylation sites,14 N-glycosylation sites and no O-glycosylation sites.Its average antigenic propensity index was 1.0340,with a total of 25 antigenic determinant cluster regions,7 B-

关 键 词：刚地弓形虫 ROP16蛋白 生物信息学 免疫原性 

分 类 号：R382.5[医药卫生—医学寄生虫学]