细粒棘球绦虫葡萄糖转运蛋白GLUT3的生物信息学分析及系统发育树

Bioinformatic analysis of and construction of a phylogenetic tree for glucose transporter type 3 of Echinococcus granulosus

目的基因克隆及获取细粒棘球绦虫葡萄糖转运蛋白3(Echinococcus granulosus Glucose transporter 3,EgGLUT3)基因全长序列,并进行生物信息学分析。方法拟通过全长基因PCR方法克隆获取EgGLUT3基因序列。利用生物信息学工具对其基因进行生物信息分析。结果从细粒棘球蚴原头蚴(E.granulosus protoscoleces,EgPSCs)成功克隆获得1 530bp的EgGLUT3基因,属于SLC2A (Solute carrier family 2)家族的GLUTs家族,相对分子量为55192.22,18-19之间有一个裂解位点。有3个酰胺化位点,7个糖基化位点,1个cAMP和cGMP依赖的蛋白激酶磷酸化位点,4个酩蛋白激酶Ⅱ磷酸化位点,11个酰基化位点,3个蛋白激酶c磷酸化位点。具有12个跨膜区域,亲水性平均值范围是-2.567~3.44。二级结构中α螺旋约占53.83%,β折叠约占16.90%,β转角约占5.30%,无规卷曲约占23.97%。从8~286氨基酸位置发现MFS家族(Major Facilitator Super family)结构域(E-value 2.5e-17)。EgGLUT3氨基酸序列与其他物种的相似性11.31%到84.82%。其基因跟人类、狗、小鼠、白马鱼和果蝇等哺乳动物的亲缘关系较远。结论这项研究为研究EgGLUT3参与E.granulosus从中间宿主体内摄取葡萄糖的机制和鉴定抗包虫药物靶点的重要组成部分,为研发新型抗包虫药物奠定基础。

Objective To clone and obtain bioinformatic information on Echinococcus granulosus glucose transporter 3(EgGLUT3). Methods full-length genomic PCR was used to clone the EgGLUT1 gene sequence,and bioinformatics tools were used to bioinformatically analyze thoses genes. Results Gene cloning and functional identification:The 1 530 bp EgGLUT3 gene was successfully cloned fromE.granulosus protoscoleces(EgPSCs),and it belongs to the GLUTs family of the SLC2 A(solute carrier family 2)family.Its molecular weight was 55 192.22 Da.The cleavage site of EgGLUT3 is between amino acids 18-19.There were 3 amidation sites,7 N-glycosylation sites,1 cAMP and cGMP-dependent protein kinase phosphorylation sites,4 casein kinase II phosphorylation sites,11 N-myristoylation sites,3 protein kinase C(PKC)phosphorylation sites,and 12 transmembrane regions in EgGLUT3.The average hydrophilicity range of EgGLUT3 was between-2.567 and 3.44.αhelices accounted for 53.83% of its secondary structure,β-sheets accounted for 16.90%,β-turns accounted for 5.30%,and random coils accounted for 23.97%.The similarity of homologous genes of EgGLUT3 is 25.85-84.82% EgGLUT3 is quite similar to Clonorchis sinensis(43.47%),and its similarity species was about 11.31 to 22.09%.A conserved major facilitator super family(MFS)(E-value 1.9 e-65)was found at amino acid 8 to 286 of EgGLUT3.The phylogenetic tree of EgGLUT3 and related genes of various species indicated that EgGLUT3 and E.multilocularis GLUT3 are highly similar and that EgGLUT3 is distantly related to glucose transporters in mammals such as humans and mice,zebrafish,and fruit flies. Conclusion This research is an important part of the study of EgGLUT3’s involvement in the glucose uptake of E.granulosus from the intermediate host and identification of targets of anti-hydatid drugs.These results have laid the foundation for the development of new anti-hydatid drugs.