DOMAIN MAPPING HIV VIF COMPLEXES BY LIMITED PROTEOLYSIS AND MASS-SPECTROMETRY

通过有限的蛋白水解和质谱法绘制 HIV VIF 复合物的结构域图

• 批准号: 8363838
• 负责人: John D Gross
• 金额: $ 0.47万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-06-01 至 2012-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8363838
• 关键词: Binding; Complex; Crystallization; Disease; Enzymes; Family; Family member; Funding; Future; Grant; HIV; Mass Spectrum Analysis; National Center for Research Resources; Primate Retrovirus; Principal Investigator; Proteins; Proteolysis; Recombinant Proteins; Recombinants; Research; Research Infrastructure; Resources; Source; Structure; Ubiquitin; United States National Institutes of Health; Viral; Viral Genome; X-Ray Crystallography; antiretroviral therapy; combat; cost; design and construction; domain mapping; reconstitution; research study; success; ubiquitin ligase

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. The APOBEC3 family of enzymes protects primates from retroviruses such as HIV by hyperediting the viral genome thereby restricting viral spread in the host. HIV Vif combats this defense by binding APOBEC3 family members and hijacking a cellular complex that catalyzes the addition of the small protein Ubiquitin to APOBEC3, sentencing the APOBEC3 proteins for destruction and allowing viral spread. We have recently reconstituted the VIF Ubiquitin ligase from recombinant proteins and seek to solve the structure of the hijacked complex by X-ray crystallography. Critical to the success of this project is characterization of the disordered regions of recombinant Vif complexes using limited proteolysis in conjunction with mass-spectrometry. The information provided by these experiments will help design constructs for crystallization trials and allow determination of the structure of the Vif/host complexes, providing useful information for future antiretroviral therapies. A

这个子项目是许多利用资源的研究子项目之一 由NIH/NCRR资助的中心拨款提供。子项目的主要支持 子项目的主要研究者可能是由其他来源提供的， 包括其它NIH来源。 列出的子项目总成本可能 代表子项目使用的中心基础设施的估计数量， 而不是由NCRR赠款提供给子项目或子项目工作人员的直接资金。 APOBEC 3酶家族通过使病毒基因组超重组从而限制病毒在宿主中的传播来保护灵长类动物免受逆转录病毒如HIV的侵害。HIV Vif通过结合APOBEC 3家族成员并劫持催化将小蛋白泛素添加到APOBEC 3的细胞复合物来对抗这种防御，使APOBEC 3蛋白被破坏并允许病毒传播。 我们最近重组的VIF泛素连接酶从重组蛋白质，并寻求解决劫持复杂的X-射线晶体学的结构。 该项目成功的关键是重组Vif复合物的无序区域的特性，结合质谱使用有限的蛋白水解。 这些实验提供的信息将有助于设计结晶试验的结构，并允许确定Vif/宿主复合物的结构，为未来的抗逆转录病毒疗法提供有用的信息。 一