中国药物警戒 ›› 2024, Vol. 21 ›› Issue (1): 40-44.
DOI: 10.19803/j.1672-8629.20230463

• 呼吸道病毒性疾病与中医药防治机制研究专栏 • 上一篇    下一篇

基于蛋白质芯片技术筛选与甲型流感病毒NS1蛋白互作的宿主因子及通路分析

张宇1, 张薇2,3, 高双荣1, 陈梦苹1, 王雅欣1, 徐英莉1, 曹姗1, 赵荣华1, 包蕾1, 李舒冉1, 孙静1, 鲍岩岩1, 耿子涵1, 郭姗姗1,*, 冀祖恩2,3#, 崔晓兰1   

  1. 1中国中医科学院中药研究所生物安全实验室,北京 100700;
    2新疆全安药业股份有限公司,新疆 库尔勒 841000;
    3新疆甘草及制品研究重点实验室,新疆 库尔勒 841000
  • 收稿日期:2023-08-01 出版日期:2024-01-15 发布日期:2024-01-18
  • 通讯作者: *郭姗姗,女,博士,中药防治病毒性疾病药效机制研究。E-mail：ssguo@icmm.ac.cn; #为共同通信作者。
  • 作者简介:张宇,女,硕士,中药防治病毒性疾病药效机制研究。
  • 基金资助:
    国家自然科学基金资助项目（82151210、81773977）

Screening and analysis of host factors interacting with NS1 proteins of influenza A virus based on the proteome chip

ZHANG Yu1, ZHANG Wei2,3, GAO Shuangrong1, CHEN Mengping1, WANG Yaxin1, XU Yingli1, CAO Shan1, ZHAO Ronghua1, BAO Lei1, LI Shuran1, SUN Jing1, BAO Yanyan1, GENG Zihan1, GUO Shanshan1,*, JI Zuen2,3#, CUI Xiaolan1   

  1. 1Biosecurity Laboratory, Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China;
    2Xinjiang Quan’an Pharmaceutical Company Limited, Korla Xinjiang 841000, China;
    3Xinjiang Key Laboratory for Research of Licorice and Products, Korla Xinjiang 841000, China
  • Received:2023-08-01 Online:2024-01-15 Published:2024-01-18

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摘要： 目的 筛选出与流感病毒Non-structural protein 1（NS1）蛋白结合的人类宿主蛋白并加以分析,确定这些结合蛋白富集的方向及关键蛋白,为抗流感病毒的新药研发提供思路。方法 将NS1样本、Biotin样本分别与HuProt™人类蛋白质组芯片进行杂交孵育,以两重复均满足 Z-Score≥3为筛选条件对与NS1蛋白有结合的宿主蛋白进行筛选得到特异性检出蛋白,实验组（NS1蛋白）与对照组（Biotin）比值I Mean_Ratio≥1.4为条件筛选出显著特异性检出蛋白。用检出的195个蛋白进行GO（Biological Process,Molecular Function,Cellular Component）和KEGG_PATHWAY 分析,通过蛋白-蛋白相互作用（PPI）及MCODE分析得到关键蛋白。结果 获得显著特异性检出蛋白195个,GO分析结果显示这些蛋白主要参与了mRNA加工、RNA结合、蛋白结合,KEGG分析主要富集到RNA降解、氨基酸的生物合成等通路。得到的4个关键蛋白DDX6、HSPD1、PKLR、MTHFD1中DDX6与RNA的合成、翻译等过程相关,而NS1蛋白可以通过调控流感病毒RNA和宿主RNA促进病毒的感染,推测DDX6可能在该过程发挥作用;其他3个蛋白目前虽然没有明确的研究指明其与流感病毒有关系,但是能在其他RNA病毒的感染过程中发挥作用。结论 与NS1结合的人类蛋白主要富集到RNA合成、加工、转录等过程中,MCODE分析得到的关键蛋白有潜力成为抗流感病毒新的靶点,但作用机制需要后续实验进行进一步验证。

关键词: 甲型流感病毒, NS1蛋白, 蛋白质芯片, 生信分析, 宿主因子, 通路

Abstract: Objective The human proteins interacting with influenza A virus (IAV) NS1 protein were obtained and analyzed to confirm the affected host biological processes and the core human proteins in the course of infection, providing thoughts for the development of drugs against influenza A virus. Methods NS1 sample and Biotin sample were hybridized with HuProt™ human proteome chip respectively, and the host protein bound to NS1 protein was screened on condition that both of repeated trials are matching Z-Score ≥ 3. The significantly specific proteins were further selected using a strict criteria (IMean_Ratio ≥ 1.4). Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) signaling pathway analysis were performed to evaluate these targets and MCODE was performed to explore the core proteins. Results A total of 195 significant specific proteins were obtained. Analysis results showed that these proteins were mainly involved in mRNA processing, RNA binding and protein binding, RNA degradation and amino acid biosynthesis. Among the four core proteins DDX6, HSPD1, PKLR and MTHFD1, DDX6 is related to RNA synthesis and translation. NS1 protein can promote viral infection by regulating IAV and host RNA, suggesting that DDX6 may be indispensable in this process. Although the other three proteins not clearly linked to influenza viruses, they can play a crucial role in the infection of other RNA viruses. Conclusion The human proteins bound to NS1 are mainly enriched in the process of RNA synthesis, processing, transcription, etc. The core proteins have potential to become new targets for anti-IAV, which need to be further verified by subsequent experiments.

Key words: influenza A virus, NS1 protein, proteome chip, bioinformatics, host factors, pathway

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