PLoS Pathog:解析脊髓灰质炎病毒2C ATPase的晶体结构,为揭示小RNA病毒科复制机制再迈进一步

2018-10-06 Tony2012 病毒学界

近日,中国医学科学院,病原生物学研究所崔胜研究员团队解析了2.55埃分辨率的脊髓灰质炎病毒(Poliovirus)2C ATPase晶体结构。

近日,中国医学科学院,病原生物学研究所崔胜研究员团队解析了2.55埃分辨率的脊髓灰质炎病毒(Poliovirus)2C ATPase晶体结构。他们发现,PV 2C与EV71 2C的结构相似。PV 2C同样通过C端alpha螺旋与邻近的2C分子结合,发生自身寡聚化。他们还发现,PV 2C具有CCCC型锌指,EV71 2C具有CCC型锌指;这类锌指的结构与以往发现的8类锌指家族均不相同,属于新类型锌指,命名为“Enterovirus 2C-like group”;PV 2C和EV71 2C锌指结构的差异决定了2C蛋白的特异性。综合近20年2C蛋白的研究成果,他们在2C六聚体上绘制了2C功能及抑制剂结合的完整图谱,为理解2C在病毒复制中的作用及2C抑制剂设计提供了关键信息。相关研究成果在线发表于PLOS Pathogens。

研究背景

自世界卫生组织发起脊髓灰质炎根除计划以来,根除PV病毒的目标仍未完成。至今,仍有少数国家存在PV病毒流行,其中2个与我国接壤(巴基斯坦和阿富汗)。为了彻底消除PV的威胁,研发抗病毒药物是重要的疾病控制手段之一。

PV属于肠道病毒C种,包括三个血清型。虽然不同的肠道病毒的衣壳存在较大差异,但其编码的复制酶高度保守。在肠道病毒编码的复制酶中,2C蛋白可能是最重要的direct-acting antivirals (DAA)药物靶标。目前,至少发现了10种2C抑制剂,包括了guanidine hydrochloride, HBB, TBZE-029, MRL-1237, pirlindole, dibucaine, zuclopenthixol, hydantoin, fluoxetine 和brefeldin A等,其中大多数抑制剂的靶点是PV 2C。然而,PV 2C的高分辨率结构信息始终无法解析,阻碍了抑制剂的研发。

结果速览

崔胜研究团队成功解析了一个PV 2C可溶性片段晶体结构。该片段缺失N-端膜结合区,但保留了完整的ATP酶结构域,锌指和C-端alpha螺旋域。他们发现,PV 2C通过C-端螺旋发生自身寡聚化(图1)。这种特征与EV71 2C高度相似,从而表明:2C蛋白通过C端两亲性helix介导六聚体组装的机制在肠道病毒中是保守的。


Fig 1. PV 2C形成依赖C-端的多聚体

PV 2C和EV71 2C在结构上最大的差异来自于锌指。EV71 2C具有一个CCC三锌指几何结构,而PV 2C具有一个CCCC四锌指几何结构。通过三维结构的比对发现,PV 2C和EV71 2C的锌指与蛋白质数据库中已知的8类锌指的构象截然不同。因此,他们将这一类新型锌指归为第9类锌指,并命名为“Enterovirus 2C-like”锌指家族(图 2 )。锌指通常参与蛋白-蛋白间相互作用。PV 2C和EV71 2C之间锌指的差异暗示了这两种蛋白特异性的差别。


Fig 2:肠道病毒2C具有新型锌指折叠,与已知的8类锌指家族均不相同

结 语

自上世纪90年代以来,人们对PV 2C蛋白进行了大量系统的研究。无数遗传学和生物化学的研究数据鉴定出PV 2C中大量的药物耐受位点和功能相关区域。崔胜研究团队把所有重要的氨基酸位点和区域首次集中展示在PV 2C六聚体模型的表面(图3)。可以看到,(1)药物耐受的氨基酸位点集中在六聚体细胞质一侧(正面),围绕在六聚体中央隧道的周围(红色和紫色区域),表明这一区域是药物作用的重要靶点;(2)锌指(黄色)位于六聚体模型的六个顶点,暗示这里是与其他蛋白相互作用的重要位置。这些发现为设计抗病毒药物提供了重要科学依据。


Fig 3:小RNA病毒2C功能及抑制剂结合位点图谱

中国医学科学院,病原生物学研究所关洪鑫(现在单位:福建师范大学生命科学学院),田娟和张楚为本文共同第一作者;中国医学科学院,病原生物学研究所崔胜研究员为本文通讯作者。该项研究得到了国家重点研究和发展计划和国家自然科学基金等资助。

原始出处:Guan H, Tian J, Zhang C, et al. Crystal structure of a soluble fragment of poliovirus 2CATPase. PLoS Pathog. 2018 Sep 19

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    2019-09-01 hongbochen
  2. 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    2019-03-18 循证小兵
  3. 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  4. [GetPortalCommentsPageByObjectIdResponse(id=1672898, encodeId=98bc16e289864, content=<a href='/topic/show?id=b79b910345c' target=_blank style='color:#2F92EE;'>#解析#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=24, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=91034, encryptionId=b79b910345c, topicName=解析)], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=f42d26916490, createdName=hongbochen, createdTime=Sun Sep 01 00:53:00 CST 2019, time=2019-09-01, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1715700, encodeId=7ad41e1570020, content=<a href='/topic/show?id=8d0f1e691b5' target=_blank style='color:#2F92EE;'>#tPA#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=32, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=17691, encryptionId=8d0f1e691b5, topicName=tPA)], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=6d2632003894, createdName=循证小兵, createdTime=Mon Mar 18 22:53:00 CST 2019, time=2019-03-18, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1941729, encodeId=c35d1941e29d4, content=<a href='/topic/show?id=c9048462146' target=_blank style='color:#2F92EE;'>#脊髓灰质炎病毒#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=34, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=84621, encryptionId=c9048462146, topicName=脊髓灰质炎病毒)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=d39f177, createdName=lixiao3326, createdTime=Mon Dec 03 21:53:00 CST 2018, time=2018-12-03, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=2024107, encodeId=a87d202410efe, content=<a href='/topic/show?id=d68b29e03d' target=_blank style='color:#2F92EE;'>#ATPase#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=34, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=2970, encryptionId=d68b29e03d, topicName=ATPase)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=f984116, createdName=kzlchina, createdTime=Sun Feb 17 14:53:00 CST 2019, time=2019-02-17, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1461669, encodeId=9736146166915, content=<a href='/topic/show?id=6f3829619b' target=_blank style='color:#2F92EE;'>#ATP#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=0, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=2961, encryptionId=6f3829619b, topicName=ATP)], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=d5806435870, createdName=listen320, createdTime=Sun Oct 07 15:53:00 CST 2018, time=2018-10-07, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1483135, encodeId=20141483135ec, content=<a href='/topic/show?id=19398461920' target=_blank style='color:#2F92EE;'>#脊髓灰质炎#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=44, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=84619, encryptionId=19398461920, topicName=脊髓灰质炎)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=4c37265, createdName=zhaojie88, createdTime=Sun Oct 07 15:53:00 CST 2018, time=2018-10-07, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1604221, encodeId=6ba6160422178, content=<a href='/topic/show?id=a2df4684e9a' target=_blank style='color:#2F92EE;'>#小RNA#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=44, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=46847, encryptionId=a2df4684e9a, topicName=小RNA)], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=2c9119152195, createdName=花花1372, createdTime=Sun Oct 07 15:53:00 CST 2018, time=2018-10-07, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1625790, encodeId=83161625e901f, content=<a href='/topic/show?id=ca8c1556656' target=_blank style='color:#2F92EE;'>#RNA病毒#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=28, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=15566, encryptionId=ca8c1556656, topicName=RNA病毒)], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=845320898943, createdName=appleandpeer, createdTime=Sun Oct 07 15:53:00 CST 2018, time=2018-10-07, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=348152, encodeId=031f3481521d, content=谢谢分享,学习了, beContent=null, objectType=article, channel=null, level=null, likeNumber=69, replyNumber=0, topicName=null, topicId=null, topicList=[], attachment=null, authenticateStatus=null, createdAvatar=https://wx.qlogo.cn/mmopen/NUyjXTCJjo6Nx0VGA4QMDzOP8oAgnV4h51EjSfehJ9QxyiaSmw8PB56BhY8KXFSxHCibXrzcy4abrzSibXQPxJ9kzrLcJyhhB8a/0, createdBy=1d3b1672603, createdName=明月清辉, createdTime=Sat Oct 06 07:10:40 CST 2018, time=2018-10-06, status=1, ipAttribution=)]
    2019-02-17 kzlchina
  5. 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    2018-10-07 listen320
  6. 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  7. 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    2018-10-07 花花1372
  8. 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  9. 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    2018-10-06 明月清辉

    谢谢分享,学习了

    0

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国家食品药品监管总局批准了北京北生研生物制品有限公司(以下简称北生研公司)的Sabin株脊髓灰质炎灭活疫苗(以下简称IPV疫苗))生产注册申请。这是继中国医学科学院医学生物学研究所的脊髓灰质炎灭活疫苗之后,我国企业自主研发的又一个新的脊髓灰质炎灭活疫苗产品。脊髓灰质炎是一种病毒引起的极具感染性的疾病,主要影响幼童。病毒通过受污染的食物和水传播, 在肠道内繁殖,再侵袭神经系统。在少数情况下,该病可造

Lancet infect dis:在孟加拉国停用口服三价脊髓灰质病毒疫苗后脊髓灰质炎的传播情况。

2016年4月起,在全球范围内,三价口服脊髓灰质炎疫苗(tOPV)被二价I型和3型口服脊髓灰质炎疫苗(bOPV)和一剂量的灭活脊髓灰质炎病毒(IPV)所取代。对于这一改变后II型脊髓灰质炎病毒或Sabin2病毒(脊髓灰质炎病毒的一种)对于II型脊髓灰质炎的再次引入或发生的传播风险尚未完全清楚。

PLOS MED:巴基斯坦的血清型1型脊髓灰质炎发病率的危险因素和短期预测:时空分析

目前,巴基斯坦对全球消除脊髓灰质炎提供了重大挑战,巴基斯坦,在2015年报告的脊髓灰质炎中,占据了73%,并在2016年占据了54%。更好地了解导致巴基斯坦脊髓灰质炎病毒传播的危险因素和运动模式,将有助于基于证据的大规模疫苗接种计划。

疫苗引爆脊髓灰质炎 威胁全球根除工作

罕见的情况下,脊髓灰质炎疫苗中被削弱的病毒可以恢复毒性,引发疫情。 在刚果民主共和国埃博拉疫情的阴影下,另一种可怕的病毒正在蔓延:脊髓灰质炎。公共卫生专家几个月来一直在努力消灭这种病毒,但它仍在传播。目前,它已经使该国29名儿童瘫痪,6月21日,有报道称在与乌干达接壤的边界上,出现了一例病例,这加剧了人们对该病毒将横扫非洲的担忧。

脊髓灰质炎病毒的单次注射疫苗

麻省理工学院(MIT)研究人员近日开发了一种可以根除脊髓灰质炎新型纳米粒子疫苗。这种疫苗只需一次注射即可提供多剂量疫苗,可以更容易地为巴基斯坦及其他发现该疾病的国家或偏远地区的儿童提供免疫接种