细粒棘球绦虫丙酮酸激酶生物信息学及系统发育分析

Bioinformatics analysis of Echinococcus granulosus sensu lato pyruvate kinase and establish its phylogenetic tree

目的运用生物信息学方法分析细粒棘球绦虫(Echinococcus granulosus sensu lato,Eg)丙酮酸激酶(Pyruvate kinase,PK)基因序列的结构与功能。方法通过NCBI蛋白数据库获取EgPK基因序列,利用ProtParam、ProtScale、SignalP 5.0、TMHMM、Motif Scan、ProtComp 9.0、CDD数据库、SOPMA、DNAStar、Swiss-Model Verify 3D、DNAMAN、VMD和MEGA等工具预测分析EgPK基因的相关生物学信息,并构建系统进化树,进行系统发育分析。结果生物信息学预测EgPK具有亲水性,分子质量62.05622 ku,无信号肽裂解位点,无跨膜区域。该蛋白含有3个N-糖基化位点,6个酪蛋白激酶Ⅱ磷酸化位点,6个N-肉豆蔻酰化位点,9个蛋白激酶C磷酸化位点。3D结构分析与序列比对显示,EgPK的桶状结构域和盖结构域之间存在特征性催化位点以及丝氨酸、苏氨酸、酪氨酸磷酸化位点。EgPK基因序列与多房棘球绦虫PK基因相似性为94.67%,与人的PK基因相似性为54.77%,EgPK与多房棘球绦虫PK蛋白亲缘关系较近,与人、小鼠、家兔等哺乳动物亲缘关系较远。结论EgPK具有PK的特征性结构域,可能参与细粒棘球蚴能量代谢相关活动。RNDFKEDT为EgPK的特征性活性位点,可作为开发抗囊型棘球蚴病的药物的候选靶点。

Objective To analyze the structure and function of the pyruvate kinase gene sequence of E.granulosus sensu lato by bioinformatics method.Methods The sequence of EgPK gene was obtained from NCBI protein database,Protparam,ProtScale,SignalP 5.0,TMHMM,Motif Scan,ProtComp 9.0,CDD database,SOPMA,DNASTAR,Swiss-Model Verify 3 D,DNAMAN,VMD,and MEGA were used to predict and analyze the biological information of EgPK gene.Results Bioinformatics predicted that EgPK was hydrophilic with a relative molecular weight of 62.05622 ku,no signal peptide cleavage site and no transmembrane region,the protein contained 3 N-glycosylation sites,6 casein kinaseⅡphosphorylation sites,6 N-nutmeg acylation sites,and 9 protein kinase C phosphorylation sites.The 3 D structural analysis and sequence alignment showed that there were characteristic catalytic sites and serine,threonine and tyrosine phosphorylation sites between the barrel-like domain and the cap domain of EgPK.The sequence of the EgPK gene was 94.67%similar to E.multilocularis and 54.77%similar to humans,it is closely related to E.multilocularis,but far from the evolutionary relationship with humans,mice,rabbits,and other mammals.Conclusion EgPK has the characteristic domain of PK and may be involved in energy metabolism of E.granulosus.RNDFKEDT is a characteristic active site of EgPK,which can be used as a candidate target to develop drugs against cystic echinococcosis.