SARS-CoV-2 Nsp16蛋白分子特征及其对男性生殖功能潜在影响

张凌寒; 张婉玉; 徐小璐; 郭艺红

doi:10.16462/j.cnki.zhjbkz.2021.04.013

SARS-CoV-2 Nsp16蛋白分子特征及其对男性生殖功能潜在影响

doi: 10.16462/j.cnki.zhjbkz.2021.04.013

450001 郑州，郑州大学第一附属医院生殖医学中心，河南省生殖与遗传重点实验室

基金项目:

国家自然科学基金 81571409

河南省高校科技创新团队支持计划 18IRTSTHN030

详细信息

通讯作者:
郭艺红，E-mail: 13613863710@163.com

中图分类号: R373
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出版历程
- 收稿日期: 2020-11-09
- 修回日期: 2021-01-26
- 网络出版日期: 2021-05-11
- 刊出日期: 2021-04-10

Molecular characteristics of SARS-CoV-2 Nsp16 protein and analysis of its potential effect on male reproductive function

Center for Reproductive Medicine, the First Affiliated Hospital of Zhengzhou University, Henan Province Key Laboratory of Reproduction and Genetics, Zhengzhou 450001, China

Funds:

National Natural Science Foundation of China 81571409

Henan Innovation Team Support Program for Colleges and Universities 18IRTSTHN030

More Information

Corresponding author: GUO Yi-hong, E-mail: 13613863710@163.com

摘要

摘要: 目的 COVID-19可能并发生殖功能损伤的问题已引起关注。本研究分析SARS-CoV-2 Nsp16蛋白遗传特征、分子结构与生物功能，探讨病毒侵入睾丸组织后Nsp16对生殖细胞的潜在影响，为该病发病机制和治疗策略研究奠定基础。方法应用生物信息技术和国际生物数据库，分析nsp16基因变异性、Nsp16空间结构与功能及对生殖细胞的潜在影响，并利用DrugBank数据库筛选可靶向结合Nsp16的药物。结果基于3种30株冠状病毒的nsp16序列构建了进化树；SARS-CoV-2毒株间nsp16基因保守性为99%；Nsp16属于亲水蛋白，在体外细胞的半衰期(half-life)是1.9 h；Nsp16具有甲基转移酶活性，具有调节精子和睾丸间质细胞基因和功能蛋白甲基化潜能；Nsp16具有线性B细胞和CTL细胞抗原表位，可能通过激发免疫反应损伤睾丸组织；从DrugBank数据库筛选出2种可靶向结合Nsp16的抑制性药物。结论 SARS-CoV-2 Nsp16是基因高度保守的功能蛋白；病毒经血管紧张素转换酶2(angiotensin-converting enzyme 2, ACE2)受体侵入睾丸组织后，Nsp16可能通过促进宿主细胞基因和蛋白甲基化机制，影响生殖细胞生长发育。该研究首次报道靶向结合Nsp16的化疗药物，对COVID-19及相关男性生殖系统疾病的防治研究具有重要参考价值。
- 核糖甲基转移酶 /
- 新型冠状病毒 /
- Nsp16 /
- 治疗策略 /
- 遗传特征
Abstract: Objective The possibility of coronavirus disease 2019 (COVID-19) involving injury to reproductive function has attracted attention. This study analyzed the genetic characteristics, molecular structure and biological function of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) Nsp16 protein, and explored potential effects of Nsp16 on germ cells following the virus′ invading testicular tissue, aiming to lay basis for studies of pathogenic mechanisms and therapeutic strategies. Methods Bioinformatic techniques and international biological databases were used to analyze nsp16 genetic variability, Nsp16 spatial structure and function, and effects on genes and proteins of germ cells. DrugBank databases were applied in screening for drugs targeted at Nsp16. Results An evolutionary tree was constructed based on the nsp16 sequences of 30 isolates of 3 coronavirus species. The nsp16 conserved property was 99% amongst SARS-CoV-2 isolates. Nsp16 is a hydrophilic protein, with a 1.9 h half-life inside cells in vitro. Nsp16 has methyltransferase activity, showing potential to regulate gene and functional protein methylation of sperm and Leydig cells. Nsp16 has both linear B cell epitopes and CTL cell epitopes, with capacity to induce immune responses and damage to testicular tissue. Two inhibitory drugs targeted at Nsp16 were found by screening the DrugBank database. Conclusions SARS-CoV-2 Nsp16 is a functional protein encoded by a highly conserved gene, may affect germ cell growth and development by promoting methylation of host cellular genes and proteins following the virus′ invasion into testis tissue through angiotensin-converting enzyme 2 receptors. This report presents Nsp16-targeted chemotherapeutic drugs for the first time, showing high reference value for prevention and treatment of COVID-19 and related lesions of the male reproductive system.
- Ribose methyltransferase /
- SARS-CoV-2 /
- Nsp16 /
- Therapeutic strategy /
- Genetic characteristic

HTML全文

图 1 SARS-CoV-2、SARS及MERS病毒株nsp16基因遗传进化树

Figure 1. Genetic evolutionary tree of nsp16 genes of SARS-CoV-2, SARS and MERS isolates

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图 2 Nsp16磷酸化位点

Figure 2. Nsp16 phosphorylation sites

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图 3 Nsp16空间结构分析

Figure 3. Analysis of spatial structure of Nsp16

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表 1 研究应用的生物软件和分析平台

Table 1. Biological softwares and analysis platforms applied in this study

生物软件/在线分析平台	URL	分析
Omiga 2.0	http://www.oxmol.co.uk/prods/omiga/	nsp16基因序列保守性
MEGA 5.10	https://www.megasoftware.net/index.php	nsp16基因进化树
Protparam	https://web.expasy.org/protparam/	Nsp16分子质量、氨基酸构成、等电点、半衰期、稳定性等理化特性
Protscale	https://web.expasy.org/protscale/	Nsp16亲疏水性
SignalP 4.0	http://www.cbs.dtu.dk/services/SignalP-4.0/	Nsp16信号肽序列
TMPRED	http://embnet.vitalit.ch/ software/TMPRED/	Nsp16跨膜螺旋
NetPhos 3.1	http://www.cbs.dtu.dk/services/NetPhos/	Nsp16磷酸化位点
NetOGlyc 4.0	http://www.cbs.dtu.dk/services/NetOGlyc-4.0/	Nsp16 O-糖基化位点
SUMOplot	http://www.abgent.com/sumoplot	Nsp16 SUMO修饰位点
CDD模块	https://www.ncbi.nlm.nih.gov/cdd/	Nsp16保守功能结构域
SOPMA	https://npsa-prabi.ibcp.fr/cgi-bin/npsa_automat.pl?page=npsa_sopma.html	Nsp16二级结构
SWISS-MODEL	https://swissmodel.expasy.org/	Nsp16三级结构
BLEP 2.0	http://tools.immuneepitope.org/bcell/	B细胞相关抗原表位
SYFPEITHI	http://www.syfpeithi.de/bin/mhcserver.dll/epitopeprediction.htm	CTL细胞相关抗原表位
DrugBank	http://www.drugbank.ca/	筛选可与Nsp16靶向结合的潜在药物分子

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