重组肠道病毒71型类病毒颗粒三维结构的测定与分析
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摘要
目的:通过测定汉逊酵母重组表达的EV71类病毒颗粒的三维结构,对重组EV71 VLP进行结构确认;通过EV71成熟病毒颗粒、天然空心颗粒及重组类病毒颗粒三者的三维结构比对,分析重组EV71 VLP与天然EV71病毒颗粒的结构异同,为重组EV71类病毒颗粒疫苗的抗原性、免疫原性及保护机制提供结构学分析依据。结果:通过单颗粒三维重构计算获得了分辨率达到近原子分辨率的EV71 VLP三维重构密度图,并根据密度图搭建了重组EV71VLP的原子模型。重组EV71类病毒颗粒是具有二十面体对称性的球状颗粒,直径33nm,在二次轴部位具有一条狭长的孔洞,连通颗粒内外,这一结构特征与EV71天然空心颗粒是一致的,而与表面完全闭合的EV71成熟病毒颗粒不同。将EV71类病毒颗粒的结构单元(protomer,由VP0、VP1与VP3三个蛋白组成)分别与EV71两种天然颗粒形式(实心、空心)各自的结构单元进行结构比对可以发现,EV71类病毒颗粒与天然空心颗粒结构单元的三维结构高度近似,而与实心颗粒(成熟病毒颗粒)存在差异,差异主要体现在VP0与VP1相对于VP3的逆时针扭转。结论:重组EV71VLP的颗粒整体特征及结构单元三维结构虽然与EV71成熟病毒颗粒存在一定的差异,但它与EV71天然空心颗粒在三维结构上高度近似,因此重组EV71 VLP在分子水平上很好的模拟了天然病毒的一种颗粒形式。
引文
[1]程爱萍.儿童传染病临床流行病学特征分析研究[J].中国儿童保健杂志,2012,20(12):1 130-32.
[2]LIU S L,PAN H,LIU P,et al.Comparative epidemiology and virology of fatal and nonfatal cases of hand,foot and mouth disease in mainland China from 2008 to 2014[J].Reviews in medical virology,2015,25(2):115-28.
[3]CHONG P,HSIEH S Y,LIU C C,et al.Production of EV71 vaccine candidates[J].Human vaccines&immunotherapeutics,2012,8(12):1775-83.
[4]MCMINN P C.An overview of the evolution of enterovirus 71 and its clinical and public health significance[J].FEMS microbiology reviews,2002,26(1):91-107.
[5]BASAVAPPA R,SYED R,FLORE O,et al.Role and mechanism of the maturation cleavage of VPO in poliovirus assembly:stracture of the empty capsid assembly intermediate at 2.9 Aresolution[J].Protein science:a publication of the Protein Society,1994,3(10):1651-69.
[6]LI H Y,HAN J F,QIN C F,et al.Virus-like particles for enterovirus 71 produced from Saccharomyces cerevisiae potently elicits protective immune responses in mice[J].Vaccine,2013,31(32):3281-7.
[7]CHUNG Y C,HO M S,WU J C,et al.Immunization with virus-like particles of enterovirus 71 elicits potent immune responses and protects mice against lethal challenge[J].Vaccine,2008,26(15):1855-62.
[8]WANG X,PENG W,REN J,et al.A sensor-adaptor mechanism for enterovirus uncoating from structures of EV71[J].Nature structural&molecular biology,2012,19(4):424-9.
[9]BLONDEL B,AKACEM O,CRAINIC R,et al.Detection by monoclonal antibodies of an antigenic determinant critical for poliovirus neutralization present on VP1 and on heat-inactivated virions[J].Virology,1983,126(2):707-10.
[10]CURRY S,FRY E,BLAKEMORE W,et al.Dissecting the roles of VPO cleavage and RNA packaging in picomavirus capsid stabilization:the stracture ofempty capsids of foot-and-mouth disease virus[J]rJournatof virology,1997,71(12):9743-52.
[11]程晨,冯敏,等.EV71两种结构形式的免疫学特性比较[J].中国生物制品学杂志,2014,27(11):1388-93.
[2]LIU S L,PAN H,LIU P,et al.Comparative epidemiology and virology of fatal and nonfatal cases of hand,foot and mouth disease in mainland China from 2008 to 2014[J].Reviews in medical virology,2015,25(2):115-28.
[3]CHONG P,HSIEH S Y,LIU C C,et al.Production of EV71 vaccine candidates[J].Human vaccines&immunotherapeutics,2012,8(12):1775-83.
[4]MCMINN P C.An overview of the evolution of enterovirus 71 and its clinical and public health significance[J].FEMS microbiology reviews,2002,26(1):91-107.
[5]BASAVAPPA R,SYED R,FLORE O,et al.Role and mechanism of the maturation cleavage of VPO in poliovirus assembly:stracture of the empty capsid assembly intermediate at 2.9 Aresolution[J].Protein science:a publication of the Protein Society,1994,3(10):1651-69.
[6]LI H Y,HAN J F,QIN C F,et al.Virus-like particles for enterovirus 71 produced from Saccharomyces cerevisiae potently elicits protective immune responses in mice[J].Vaccine,2013,31(32):3281-7.
[7]CHUNG Y C,HO M S,WU J C,et al.Immunization with virus-like particles of enterovirus 71 elicits potent immune responses and protects mice against lethal challenge[J].Vaccine,2008,26(15):1855-62.
[8]WANG X,PENG W,REN J,et al.A sensor-adaptor mechanism for enterovirus uncoating from structures of EV71[J].Nature structural&molecular biology,2012,19(4):424-9.
[9]BLONDEL B,AKACEM O,CRAINIC R,et al.Detection by monoclonal antibodies of an antigenic determinant critical for poliovirus neutralization present on VP1 and on heat-inactivated virions[J].Virology,1983,126(2):707-10.
[10]CURRY S,FRY E,BLAKEMORE W,et al.Dissecting the roles of VPO cleavage and RNA packaging in picomavirus capsid stabilization:the stracture ofempty capsids of foot-and-mouth disease virus[J]rJournatof virology,1997,71(12):9743-52.
[11]程晨,冯敏,等.EV71两种结构形式的免疫学特性比较[J].中国生物制品学杂志,2014,27(11):1388-93.