MULTIFREQUENCY STUDIES OF SPIN-LABELED HIV-1 PROTEASE

自旋标记 HIV-1 蛋白酶的多频研究

• 批准号: 8363996
• 负责人: KEITH A EARLE
• 金额: $ 0.26万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-01 至 2012-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8363996
• 关键词: Electron Spin Resonance Spectroscopy; Environment; Experimental Designs; Funding; Grant; HIV; HIV-1; Learning; Measurement; Membrane Proteins; Monitor; National Center for Research Resources; Peptide Hydrolases; Pharmaceutical Preparations; Principal Investigator; Property; Protease Inhibitor; Proteins; Research; Research Infrastructure; Resources; Site; Source; Spin Labels; Structure; Surgical Flaps; Technology; United States National Institutes of Health; base; cost; inhibitor/antagonist; interest; response; tool

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. Site-directed spin labeling (SDSL) is a powerful tool for monitoring the structure and dynamics of both soluble and membrane proteins. Measurements of the spectral properties of the paramagnetic nitroxide probe with electron paramagnetic resonance (EPR) spectroscopy provide a wealth of information on the environment in the protein. The HIV-1 protease is a 22 kDa homodimeric protein essential for function of the AIDS virus, and protease inhibitors have been developed into effective HIV drugs. We are particularly interested in the details of HIV-1 protease flap dynamics that can be learned from multifrequency SDSL EPR studies. Earlier SDSL studies of several spin labels at 9.5 GHz do not reveal any significant change in the flap dynamics in the presence or absence of protease inhibitor, contrary to expectration. Preliminary results at 170 GHz do indicate small but discernible differences in the flap dynamics in the presence or absence of inhibitor with the inhibited form showing clear evidence of slower tumbling. Based on these promising initial results we are refining the experimental design in order to enhance the spectral differences. We suggest that complementary studies at 240 GHz will further resolve the spectral sensitivity to the observed small differences in dynamics and will provide a useful means for quantifying the response of HIV-1 protease to the presence of various inhibitors.

该副本是利用资源的众多研究子项目之一 由NIH/NCRR资助的中心赠款提供。对该子弹的主要支持 而且，副投影的主要研究员可能是其他来源提供的 包括其他NIH来源。 列出的总费用可能 代表subproject使用的中心基础架构的估计量， NCRR赠款不直接向子弹或副本人员提供的直接资金。 位置定向的自旋标记（SDSL）是监测可溶性和膜蛋白的结构和动力学的强大工具。通过电子顺磁共振（EPR）光谱验证了顺磁性氮氧化物探针的光谱特性的测量，提供了有关蛋白质环境的大量信息。 HIV-1蛋白酶是22 kDa同型二聚体对于艾滋病病毒功能所必需的，并且蛋白酶抑制剂已发展为有效的HIV药物。 我们对可以从多频SDSL EPR研究中学到的HIV-1蛋白酶皮瓣动态的细节特别感兴趣。 早期对9.5 GHz的自旋标记的SDSL研究并未揭示在存在或不存在蛋白酶抑制剂的情况下，与预期相反。 在170 GHz处的初步结果确实表明在存在或不存在抑制剂的情况下，lap动力学的差异很小但可见的差异，抑制形式显示出明显的较慢的表现。 基于这些有希望的初始结果，我们正在完善实验设计，以增强光谱差异。 我们建议，在240 GHz的互补研究将进一步解决对观察到的动力学差异的频谱敏感性，并将为量化HIV-1蛋白酶对存在各种抑制剂的存在的响应提供有用的手段。