Insights into biomolecular reactivity and structure for virus inactivation prediction

深入了解病毒灭活预测的生物分子反应性和结构

• 批准号: 2212779
• 负责人: Yinyin Ye
• 金额: $ 30万
• 依托单位: SUNY at Buffalo
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-01 至 2025-08-31
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2212779&HistoricalAwards=false
• 关键词: Insights; into; biomolecular; reactivity; structure

The emergence and spread of new viruses such as SARS-CoV-2 and its variants has heightened the need to assess the efficacy of virus inactivation by disinfectants including chemical oxidants such as chlorine and chlorine dioxide. Culture-based approaches are the current gold standard methods to track the levels of infective viruses before and after disinfection. However, many viruses are not culturable, or are too dangerous to be cultured, and thus their mechanisms of inactivation by chemical oxidants are not well understood. The overarching goal of this project is to investigate and unravel the fundamental reactions and structural changes of viral proteins that occur during virus inactivation by chemical oxidants such as chlorine and chlorine dioxide. To advance this goal, the Principal Investigator (PI) propose to integrate high-throughput and high-sensitivity proteomic analysis, structural analysis, and data mining to test the hypothesis that virus inactivation by oxidants is driven by the decay of viral proteins, and the decay rate constants of viral peptides is a function of solvent accessibility of susceptible amino acid residues in the peptide sequences. The successful completion of this project will benefit society through the development of a mechanistic understanding of virus susceptibility to disinfectants that could provide guidance to the public and water utilities regarding the selection of the most efficient chemical oxidants and disinfectant dosages to inactivate waterborne viruses while minimizing the formation of toxic disinfection byproducts. Additional benefits to society will be achieved through student education and training including the mentoring of a graduate student at the University at Buffalo.A virus particle consists of a single molecule of genome, which is surrounded by a protein capsid, and/or lipid envelope. The viral genome and proteins carry various biological functions that are essential for virus infection. Previous studies of virus inactivation by UV and chemical oxidants suggest that the degradation of viral biomacromolecules, particularly the genome and proteins, correspond to the loss of virus infectivity. However, a fundamental understanding of the mechanisms of virus inactivation by chemical oxidants has remained elusive. To address this critical knowledge gap, the Principal Investigator (PI) of this project proposes to integrate high-throughput and high-sensitivity proteomic analysis, structural analysis, and data mining to investigate and unravel the mechanisms of oxidation of viral proteins by chemical disinfectants. The specific objectives of the research are to: (1) Characterize viral protein degradation by chemical oxidants and identify structural features of viral peptides that drive virus inactivation using chlorine and chlorine dioxide as model disinfectants; (2) Evaluate the impacts of oxidative modifications and peptide cleavages on the conformational change of viral proteins; and (3) Evaluate the impacts of lipid permeability to oxidant molecules on the extents and rates of inactivation of enveloped waterborne viruses. The successful completion of this research has the potential for transformative impact through the development and validation of a new model that could predict the decay kinetics of viral proteins when viruses are treated by oxidants and help identify the biomolecular features of viral proteins that control virus susceptibility or resistance to disinfectants. To implement the education and outreach activities of the project, the PI plans to incorporate the findings from this research into undergraduate and graduate courses at the University at Buffalo (UB). In addition, the PI proposes to leverage the UB Louis Stokes Alliance for Minority Participation (LSAMP) Summer Research Internship Program to recruit two undergraduate students from underrepresented groups that will work on the project.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

SARS-CoV-2及其变种等新病毒的出现和传播，使得评估消毒剂（包括氯和二氧化氯等化学氧化剂）灭活病毒的有效性变得更加必要。基于培养的方法是目前追踪消毒前后感染性病毒水平的金标准方法。然而，许多病毒是不可培养的，或者培养起来太危险，因此它们被化学氧化剂灭活的机制尚不清楚。该项目的总体目标是研究和揭示化学氧化剂（如氯和二氧化氯）灭活病毒过程中发生的病毒蛋白质的基本反应和结构变化。 为了推进这一目标，主要研究者（PI）建议整合高通量和高灵敏度蛋白质组学分析、结构分析和数据挖掘，以检验以下假设：氧化剂灭活病毒是由病毒蛋白的衰变驱动的，病毒肽的衰变速率常数是肽序列中敏感氨基酸残基的溶剂可及性的函数。该项目的成功完成将通过发展对病毒对消毒剂的敏感性的机械理解而造福社会，这可以为公共和供水设施提供指导，以选择最有效的化学氧化剂和消毒剂剂量来抑制水传播病毒，同时最大限度地减少有毒消毒副产物的形成。通过对学生的教育和培训，包括对布法罗大学研究生的指导，将为社会带来额外的好处。病毒颗粒由单个基因组分子组成，被蛋白质衣壳和/或脂质包膜包围。病毒基因组和蛋白质携带病毒感染所必需的各种生物学功能。以前的研究表明，紫外线和化学氧化剂灭活病毒的生物大分子，特别是基因组和蛋白质的降解，对应于病毒感染性的丧失。然而，对化学氧化剂灭活病毒的机制的基本理解仍然难以捉摸。为了解决这一关键的知识缺口，该项目的主要研究者（PI）建议整合高通量和高灵敏度的蛋白质组学分析，结构分析和数据挖掘，以研究和揭示化学消毒剂氧化病毒蛋白的机制。本研究的具体目的是：（1）以氯和二氧化氯为模型消毒剂，表征化学氧化剂对病毒蛋白的降解，并确定驱动病毒灭活的病毒肽的结构特征;（2）评价氧化修饰和肽裂解对病毒蛋白构象变化的影响;以及（3）评价脂质对氧化剂分子的渗透性对有包膜的水性病毒的灭活程度和速率的影响。这项研究的成功完成有可能通过开发和验证一种新的模型来产生变革性的影响，该模型可以预测病毒蛋白质在氧化剂处理时的衰变动力学，并有助于识别控制病毒易感性或耐消毒剂性的病毒蛋白质的生物分子特征。为了实施该项目的教育和推广活动，PI计划将这项研究的结果纳入布法罗大学（UB）的本科生和研究生课程。此外，PI建议利用UB Louis Stokes少数民族参与联盟（LSAMP）暑期研究实习计划，从代表性不足的群体中招募两名本科生，他们将参与该项目。该奖项反映了NSF的法定使命，并通过使用基金会的智力价值和更广泛的影响审查标准进行评估，被认为值得支持。