机构地区:[1]大理大学基础医学院医学微生物学及免疫学教研室,云南大理671000 [2]大理大学云南省昆虫生物医药研发重点实验室,云南大理671000
出 处:《微生物学杂志》2023年第5期81-90,共10页Journal of Microbiology
基 金:云南省地方本科高校联合专项面上项目(2017FH001-085);大理大学高层次人才资助项目(KYBS201708);大理大学临床分子免疫学创新团队项目(ZKLX2019105)。
摘 要:运用生物信息学分析软件预测结核分枝杆菌(Mycobacterium tuberculosis,Mtb)Rv0081蛋白的生物学特征及筛选潜在的优势抗原表位。从NCBI数据库获取Mtb Rv0081蛋白的氨基酸序列,利用生物信息学分析软件ProtParam、ProtScale及TMPRED分析Rv0081蛋白的理化性质及亲疏水性;TMHMM、SignalP-5.0 Server预测蛋白的跨膜区及信号肽;NetNGlyc-1.0 Server、NetPhos 3.1 Server分别预测蛋白的糖基化位点及磷酸化位点;STRING预测能与Rv0081相互作用的蛋白;分别运用SOPMA、SWISS-MODEL预测蛋白的二、三级结构;综合运用softberry、WoLF PSORT预测蛋白的亚细胞定位;运用DNAStar预测蛋白的B细胞抗原表位;综合运用SYFPEITHI、NetCTL 1.2 Server、Net MHC pan 4.1 server预测蛋白的CTL细胞抗原表位;综合运用SYFPEITHI、Net MHCII pan 4.0 server预测蛋白的Th细胞抗原表位。结果表明,Rv0081蛋白由114个氨基酸组成,相对分子质量为12356.32,亚细胞定位于细胞质中,为稳定的疏水性蛋白,无跨膜区和信号肽,含有1个糖基化位点及9个磷酸化位点;二级结构主要由α-螺旋和无规则卷曲构成,结构较松散;与hycE、hycP、Rv0088、Rv0083、hycD、hycQ、Rv0082、devR、Rv0080及Rv0079蛋白存在相互作用关系;综合分析各软件预测结果筛选出6个优势B细胞抗原表位、6个优势CTL细胞抗原表位及7个优势Th细胞抗原表位。Mtb Rv0081蛋白具有较多潜在的候选B、T细胞抗原表位,可作为研发新型结核疫苗的候选抗原。Biological characteristics and screening potential dominant epitopes of the Mycobacterium tuberculosis Rv0081(Mtb Rv0081)gene encoding protein was predicted using bioinformatics analysis software.Amino acid sequence of Mtb Rv0081 protein was obtained from NCBI database.The physical and chemical properties and hydrophobicity of Rv0081 protein were analyzed adopting bioinformatics analysis software ProtParam,ProtScale and TMPRED;the transmembrane regions and signal peptides of Rv0081 protein were predicted using TMHMM,SignalP-5.0 Server;glycosylation site and phosphorylation site of the Rv0081 protein were predicted by NetNGlyc-1.0 Server and NetPhos 3.1 Server,respectively;STRING was used to analyze the proteins interacting with Rv0081;the secondary and tertiary structure of the Rv0081 protein were predicted by SOPMA and SWISS-MODELr,respectively;softberry and WoLFPSORT were used to predicted the subcellular localization of the Rv0081 protein;and DNAStar,SYFPEITHI,NetCTL 1.2 Server,Net MHC pan 4.1 server and Net MHCII pan 4.0 server were used to predict the protein epitopes and then to find the dominant B and T cell epitopes.The results showed that Rv0081 protein was composed of 114 amino acids with relative molecular mass of 12356.32.The subcellular was localized at cytoplasm and was a stable hydrophobic protein without transmembrane region and signal peptide containing 1 glycosylation site and 9 phosphorylation sites.The secondary structure was mainly composed ofα-helix and random coils,and the structure was relatively loose;interacted with proteins hycE,hycP,Rv0088,Rv0083,hycD,hycQ,Rv0082,devR,Rv0080,and Rv0079;based on the comprehensive analysis of the prediction results of each software,6 dominant B cell epitopes,6 dominant CTL cell epitopes,and 7 dominant cell epitopes were screened.The Mtb Rv0081 protein containing multiple B and T cell epitopes,and can be used as a candidate protein for the development of a new vaccine against tuberculosis.
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