细粒棘球蚴DNA损伤修复蛋白RAD51的生物信息学分析及初步验证  

DNA damage repair protein RAD51 in Echinococcus granulosus bioinformatics analysis and preliminary validation

作　　者：巩月红[1,2] 林玉霞 王梦君 潘美驰 赵一聪 赵军[1,2] 王建华[1,2] GONG Yuehong;LIN Yuxia;WANG Mengjun;PAN Meichi;ZHAO Yicong;ZHAO Jun;WANG Jianhua(Pharmacy,First Hospital Affiliated with Xinjiang Medical University,Urumqi 830011,China;Xinjiang Medical University Jointly Established the State Key Laboratory of Pathogenesis,Prevention,and Treatment of Diseases Highly Prevalent in Central Asia;College of Pharmacy,Xinjiang Medical University;Medical Department,Xinjiang Medical University)

机构地区：[1]新疆医科大学第一附属医院药学部,新疆乌鲁木齐830011 [2]新疆医科大学省部共建中亚高发病成因与防治国家重点实验室 [3]新疆医科大学药学院 [4]新疆医科大学医学部

出　　处：《中国病原生物学杂志》2023年第12期1416-1423,共8页Journal of Pathogen Biology

基　　金：新疆维吾尔自治区科学技术厅自然科学基金重点项目(No.2021D01D15);省部共建国家重点实验室开放课题(No.SKLHIDCA-2022-9);新疆维吾尔自治区科学技术厅自然科学基金面上项目(No.2020D01C240)。

摘　　要：目的利用生物信息学软件预测和分析细粒棘球蚴DNA损伤修复蛋白(EgRAD51)的分子特性,为EgRAD51的功能和药物靶点研究奠定基础。方法在NCBI数据库中检索下载EgRAD51蛋白的氨基酸序列,采用ProtParam预测EgRAD51蛋白的理化性质,SignalP-5预测信号肽,ProtScale预测其亲疏水性,TMHMM分析跨膜区域,EukmPLoc2.0预测基亚细胞定位,SOMPA和Swissmodel预测其二、三级结构,NetPhos3.1、NetOGlyc4.0及NetNGlyc1.0预测磷酸化位点,ABC pred和SYFPEITHI数据库预测其T、B细胞表位。采用MEGA11软件对不同物种来源的RAD51蛋白进行多序列比对并构建系统进化树。采用qRT-PCR检测不同药物干预后EgRAD51 mRNA表达水平。结果预测EgRAD51氨基酸数378个,分子式为C1839H2950N508O544S20,分子质量为41.52193 ku,pI值为7.95;该蛋白无信号肽和无跨膜区,定位于细胞核内;含有32个丝氨酸磷酸化位点,26个苏氨酸磷酸化位点,9个酪氨酸磷酸化位点,4个O-糖基化潜在位点,1个N-糖基化位点;含PRK09302超级家族保守结构域及RAD51C重组酶保守结构域;二级结构中α-螺旋占45.24%,延伸主链占15.08%,β-转角占3.97%,无规则卷曲占35.71%,为低聚单体;含有13个B细胞抗原表位,具有能与HLA-A*02-01的结合能力,且能被分子呈递。同源性及系统进化分析显示,EgRAD51与多房棘球绦虫RAD51的氨基酸序列一致,与亚洲带绦虫、中殖孔绦虫、血吸虫、欧洲锥虫等亲缘关系较近,与智人、果蝇、中华枝睾吸虫、带翅棘球蚴亲缘关系较远。qRT-PCR检测不同药物干预组的EgRAD51 mRNA较空白对照组均上调。结论EgRAD51蛋白定位于细胞核内,含有丰富的B细胞抗原表位,具有能与HLA-A*02-01的结合能力,是一个具有潜在研究价值的药物靶点。Objective To predict and analyze the molecular characteristics of the DNA damage repair protein(EgRAD51)in Echinococcus granulosus using bioinformatics software,in order to lay the foundation for further exploring the function of EgRAD51 and developing drug targets.Methods The amino acid sequence of EgRAD51 protein was retrieved and downloaded from the NCBI database.ProtParam was used to predict the physical and chemical properties of EgRAD51 protein,SignalP-5 was used to predict the Signal peptide,ProtScale was used to predict its hydrophobicity,TMHMM was used to analyze the transmembrane region,Euk-mPLoc2.0 was used to predict the subcellular location of the protein,SOMPA and Swissmodel were used to predict its secondary and tertiary structure,NetPhos3.1,NetOGlyc4.0 and NetNGlyc1.0 were used to predict the phosphorylation site,ABC pred and SYFPEITHI databases predict the T and B cell epitopes of EgRAD51 protein.Using MEGA11 software to perform multiple sequence alignment of RAD51 proteins from different species and construct a phylogenetic tree.Using qRT PCR to detect the expression level of EgRAD51 mRNA after different drug interventions.Results The results predicted that EgRAD51 had 378 amino acid numbers,a molecular formula of C1839H2950N508O544S20,a molecular weight of 41.52193 ku,and a pI value of 7.95;The protein has no signal peptide or transmembrane region and is localized within the nucleus;Contains 32 serine phosphorylation sites,26 threonine phosphorylation sites,9 tyrosine phosphorylation sites,4 O-glycosylation potential sites,and 1 N-glycosylation site;Contains the conserved domains of PRK09302 superfamily and RAD51C recombinase;In the secondary structureα-Helixes account for 45.24%,and extended main chains account for 15.08%,β-Corners account for 3.97%,irregular curls account for 35.71%,and are oligomers;It contains 13 B cell antigen epitopes and has the ability to bind to HLA-A*02-01,and can be presented by molecules.Homology and phylogenetic analysis showed that the amino acid sequences of Eg

关 键 词：细粒棘球蚴 EgRAD51 DNA损伤修复 生物信息学 

分 类 号：R383.33[医药卫生—医学寄生虫学]