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Cell Res:突破!黄超兰与高福团队描绘新冠刺突蛋白糖基化图谱, 揭示“O-Follow-N”糖基化新规律

2021-08-04 生物探索 生物探索

此研究将深入了解新型冠状病毒(SARS-CoV-2)刺突蛋白(Spike, S)的糖基化修饰对于新型冠状病毒肺炎(COVID-19)发病机制的探索,疫苗和治疗药物的设计开发,以及检测试剂盒的生产具有重

蛋白质糖基化修饰是生物体内最重要的翻译后修饰之一,发生在细胞50%-70%的蛋白上。病毒囊膜蛋白的糖基化修饰具有广泛的功能,包括调控蛋白质稳定性、病毒的趋向性、和保护潜在的抗原表位免受免疫监视等。深入了解新型冠状病毒(SARS-CoV-2)刺突蛋白(Spike, S)的糖基化修饰对于新型冠状病毒肺炎(COVID-19)发病机制的探索,疫苗和治疗药物的设计开发,以及检测试剂盒的生产具有重要意义。此前研究者在体外纯化表达的S蛋白胞外域和从病毒颗粒中提取的S蛋白中共鉴定到了22个N-糖基化修饰位点1,2。而由于技术和样本来源的限制,已有研究仅在纯化的S蛋白上鉴定到了一些O-糖基化修饰位点,截止目前,尚未进行病毒颗粒上S蛋白的O-糖基化修饰的研究。

近日,北京大学医学部精准医疗多组学研究中心黄超兰团队,和中国科学院院士高福团队,中国科学院天津工业生物技术研究所高峰团队等开展合作研究,采用基于质谱的糖基化鉴定技术,首次揭示了病毒颗粒上提取的S蛋白O-糖基化修饰图谱,并提出了“O-Follow-N”的O糖基化修饰规律。该研究以“ O-glycosylation pattern of the SARS-CoV-2 spike protein reveals an“O-Follow-N” rule ”为题于2021年8月2日线上发表在 Cell Research 期刊上。

为获得天然状态下S蛋白的N-和O-糖基化修饰完整图谱,研究者从SARS-CoV-2病毒颗粒上获得S蛋白,用多种蛋白酶酶解成肽段,采用纳升液相色谱以及具有超高分辨率的Orbitrap Eclipse Tribrid三合一质谱联用仪,利用阶梯能量HCD(stepped collisional energy SCE),HCD(Higher-energy collisional dissociation)以及HCDpdEThcD三种碎裂方法进行质谱分析。

本研究中,研究者不但成功鉴定到了此前已报道的22个N-糖基化修饰位点,还首次从SARS-CoV-2病毒颗粒中提取的S蛋白上鉴定到了17个O-糖基化修饰位点。值得注意的是,研究者发现在这17个位点中,有11个位点位于糖基化的天冬酰胺(Asn)附近。研究者将NxS/T共有基序内糖基化的Asn每一侧的3个氨基酸定义为“N±1-3”。分析结果显示,11个O-糖基化修饰位点分布在“N±1-3”的位置上,位点信息确定的位点数有10个,其中7个位点分布在“N+2”的位置上。研究者还通过开展定点突变实验进一步证实Asn糖基化修饰的存在是“N±1-3”的位置上出现O-糖基化修饰的先决条件。综上,研究者提出SARS-CoV-2病毒S蛋白的糖基化修饰存在O-糖基化修饰追随N-糖基化修饰发生的现象,并将这一现象命名为“O-Follow-N”规律。

图. SARS-CoV-2病毒S蛋白的糖基化修饰遵循“O-Follow-N”规律

本研究基于前沿的质谱鉴定技术,揭示了S蛋白的O糖基化修饰谱,提出了O 糖基化修饰的“O-Follow-N”规律,这一规律可能适用于其它蛋白,提示O-糖基化修饰具有潜在的新机制,特别是N-和O-糖基化修饰之间可能存在的协同作用,未来有望在极大程度上推动糖生物学领域的研究。

此前,黄超兰主任领衔的多组学中心团队还与高福院士领衔的多学科团队紧密合作,揭示早期的新冠感染患者存在显着的免疫抑制,并首次提出COVID-19的发病机制或存在“两阶段”模式3。多组学中心在黄超兰教授的带领下,将继续基于临床,前沿技术和基础学科的深度交叉融合,深耕前沿技术方法开发,为推动基础生物学和临床领域的创新研究提供最有质量保证的蛋白质组和质谱技术手段。

中国科学院微生物研究所高福院士,北京大学医学部精准医疗多组学研究中心主任黄超兰教授,北京大学医学部精准医疗多组学研究中心陈扬副研究员,中国科学院天津工业生物技术研究所高峰教授为本文的共同通讯作者;北京大学医学部精准医疗多组学研究中心田文敏博士,中国科学院天津工业生物技术研究所李德林博士,北京大学医学部精准医疗多组学研究中心博士研究生张楠,中国科学院天津工业生物技术研究所博士研究生白桂杰、原恺博士为本文的共同一作。

原始出处:

Tian, W., Li, D., Zhang, N. et al. O-glycosylation pattern of the SARS-CoV-2 spike protein reveals an “O-Follow-N” rule. Cell Res (2021). https://doi.org/10.1038/s41422-021-00545-2.

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    2021-08-06 lfyang
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    2021-08-06 xsm923
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    2021-08-05 公卫新人

    新冠肺炎,疫情何时才能消失

    0

  7. [GetPortalCommentsPageByObjectIdResponse(id=1962575, encodeId=952919625e575, content=<a href='/topic/show?id=dbe6445416' target=_blank style='color:#2F92EE;'>#CEL#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=66, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=4454, encryptionId=dbe6445416, topicName=CEL)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=c3ff68, createdName=维他命, createdTime=Wed Oct 13 08:06:40 CST 2021, time=2021-10-13, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1872262, encodeId=e48618e22620a, content=<a href='/topic/show?id=fd764459a8' target=_blank style='color:#2F92EE;'>#Cell#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=66, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=4459, encryptionId=fd764459a8, topicName=Cell)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=524d95, createdName=zhaozhouchifen, createdTime=Sat Jun 11 12:06:40 CST 2022, time=2022-06-11, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=2060458, encodeId=1695206045829, content=<a href='/topic/show?id=9c5a3243ef4' target=_blank style='color:#2F92EE;'>#刺突蛋白#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=0, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=32437, encryptionId=9c5a3243ef4, topicName=刺突蛋白)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=4cce318, createdName=wleon8895, createdTime=Sun Jul 17 09:06:40 CST 2022, time=2022-07-17, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1318319, encodeId=a6ac13183190f, content=<a href='/topic/show?id=b5dde6644b1' target=_blank style='color:#2F92EE;'>#糖基化#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=83, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=76644, encryptionId=b5dde6644b1, topicName=糖基化)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=43cb463, createdName=lfyang, createdTime=Fri Aug 06 14:06:40 CST 2021, time=2021-08-06, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1574407, encodeId=ff5e15e4407f3, content=<a href='/topic/show?id=3c2c586e23f' target=_blank style='color:#2F92EE;'>#新规#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=75, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=58672, encryptionId=3c2c586e23f, topicName=新规)], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=a2ca16016783, createdName=xsm923, createdTime=Fri Aug 06 14:06:40 CST 2021, time=2021-08-06, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1029824, encodeId=046f10298242d, content=新冠肺炎,疫情何时才能消失, beContent=null, objectType=article, channel=null, level=null, likeNumber=80, replyNumber=0, topicName=null, topicId=null, topicList=[], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=f0620, createdName=公卫新人, createdTime=Thu Aug 05 02:06:40 CST 2021, time=2021-08-05, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1005238, encodeId=3c47100523879, content=蛋白质糖基化修饰是生物体内最重要的翻译后修饰之一,发生在细胞50%-70%的蛋白上。。, beContent=null, objectType=article, channel=null, level=null, likeNumber=98, replyNumber=0, topicName=null, topicId=null, topicList=[], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=bd984965877, createdName=1453df99m58暂无昵称, createdTime=Wed Aug 04 23:11:11 CST 2021, time=2021-08-04, status=1, ipAttribution=)]
    2021-08-04 1453df99m58暂无昵称

    蛋白质糖基化修饰是生物体内最重要的翻译后修饰之一,发生在细胞50%-70%的蛋白上。。

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