CAJ | 학술논문

目的分析肠道病毒71型(EV71)VP1基因及其编码蛋白的结构与功能,以指导其生物学功能研究.方法利用多种生物信息在线分析工具和分析软件包分析EV71 2008-GZCH07株VP1基因及编码蛋白与其他EV71代表株的序列,进行多序列同源比对,并构建分子进化树.预测该基因编码的蛋白质理化特性,二级结构、三维结构等结构特性,酶学特性和抗原表位等免疫学特性.结果 2008-GZCH07株与ZJ001株同源性最高(97%和98%),与人柯萨奇病毒A16同源性最低,与EV71 A、B、C型病毒的同源性为86%～98%.2008-GZCH07株与ZJ001株和 BJ08-Z025-5株的进化距离较近,属于C4亚型;A、B、C三型EV71的VP1编码蛋白中,2008-GZCH07株VP1编码蛋白与B、C型进化距离较A型近.VP1基因全长510 bp,开放读码框位于116～510 bp区域,编码132个氨基酸,等电点为4.39.VP1蛋白富含α螺旋和无规卷曲,无跨膜区域,含5个高疏水性区域,属于胞外蛋白.蛋白质同源建模分析表明该区域位于蛋白表面,形成环状结构,含有5个抗原簇,7个关键催化位点位于该区域内或其附近.结论 EV71-VP1基因编码蛋白分布有多个磷酸化位点,具有较多的生物功能位点和潜在的抗原表位区域,可能是免疫诊断、药物作用研究及疫苗研制的靶抗原,可为EV71诊断、治疗及预防方面的应用价值提供重要的参考依据.
목적 분석장도병독71형(EV71)VP1기인급기편마단백적결구여공능,이지도기생물학공능연구.방법 이용다충생물신식재선분석공구화분석연건포분석EV71 2008-GZCH07주VP1기인급편마단백여기타EV71대표주적서렬,진행다서렬동원비대,병구건분자진화수.예측해기인편마적단백질이화특성,이급결구、삼유결구등결구특성,매학특성화항원표위등면역학특성.결과 2008-GZCH07주여ZJ001주동원성최고(97%화98%),여인가살기병독A16동원성최저,여EV71 A、B、C형병독적동원성위86%～98%.2008-GZCH07주여ZJ001주화 BJ08-Z025-5주적진화거리교근,속우C4아형;A、B、C삼형EV71적VP1편마단백중,2008-GZCH07주VP1편마단백여B、C형진화거리교A형근.VP1기인전장510 bp,개방독마광위우116～510 bp구역,편마132개안기산,등전점위4.39.VP1단백부함α라선화무규권곡,무과막구역,함5개고소수성구역,속우포외단백.단백질동원건모분석표명해구역위우단백표면,형성배상결구,함유5개항원족,7개관건최화위점위우해구역내혹기부근.결론 EV71-VP1기인편마단백분포유다개린산화위점,구유교다적생물공능위점화잠재적항원표위구역,가능시면역진단、약물작용연구급역묘연제적파항원,가위EV71진단、치료급예방방면적응용개치제공중요적삼고의거.
Objective To understand the structures and functions of enterovirus 71 (EV71)VP1 gene and its encoded protein using bioinformatics method, so as to direct studies of its biological function. Methods VP1 gene and its encoded protein of EV71 2008-GZCH07 strain and other representative EV71 strains were analyzed by online analysis at bioinformatics websites and software packages. Multi-sequence homological alignment and phylogenetic analysis were performed.Physicochemical characteristics, secondary structure, homology modeling of tertiary structure,enzymological characteristics, antigenic epitope of VP1 gene encoded protein were predicted. Results The homology of EV71 2008-GZCH07 strain was highest (97% and 98%) with ZJ001 strain, and lowest with human coxsackievirus A16. The homology of EV71 2008-GZCH07 strain and EV71 types A,B,C was 86%-98%. Phylogenetic analysis demonstrated that 2008-GZCH07 stain was close to ZJ001 and BJ08-Z025-5 stains, which belonged to C4 subtype. In VP1 encoded proteins of EV71 types A,B,C, the relationship between 2008-GZCH07 and EV71-B, EV71-C was closer than EV71-A.The whole length of VP1 gene was 510 bp, with open reading frame (ORF) located at 116-510 bp region,and it encoded 132 amino acids with isoelectric point of 4.39.The protein was rich of a-helix and random coilon without transmembrane regions, and contained 5 high hydrophobic regions and belonged to extracellular protein. The homology modeling of tertiary structure showed that the region was on the surface of protein and formed a binding loop. There was 5 antigen epitopes. And 7 key catalytic sites were located at or close to the loop. Conclusions EV71-VP1 encoded protein contains many phosphorylation sites, with many biological function sites and antigenic epitope regions, which might be a potential target antigen for immunodiagnosis, anti-schistosome drug and vaccine development, and would be basis of further study of diagnosis, treatment and prevention of EV71 infection.