Nano-lmaging APOBECS Interactions

纳米成像 APOBECS 相互作用

基本信息

批准号：
8078337
负责人：
YURI L LYUBCHENKO
金额：
$ 26.75万
依托单位：
UNIVERSITY OF MINNESOTA
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-03-01 至 2015-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8078337
关键词：
AIDS/HIV problem APRIN gene Acquired Immunodeficiency Syndrome Address Area Asses Atomic Force Microscopy Binding Biochemical Chemicals Complex Core Protein DNA DNA Binding DNA Sequence Data Development Dimerization Foundations Future Genetic Goals HIV HIV therapy HIV-1 Holoenzymes Homo Human Image Imaging technology Laboratories Measurement Measures Mediating Methods Modality Molecular Molecular Analysis Nucleic Acid Binding Nucleic Acids Paper Property Proteins RNA Resolution Role Single-Stranded DNA Specificity Spectrum Analysis Structure System Technology Temperature Testing Therapeutic Viral Proteins Work design divalent metal experience imaging probe inhibitor/antagonist instrument intermolecular interaction metal chelator mutant nano nanoimaging nanoscale novel programs protein protein interaction single molecule structural biology systems research

项目摘要

APOBECSG (A3G) and APOBECSF (A3F) are involved in various anti-viral protein and nucleic acid transactions. Important primary interactions include interprotein interactions, binding single-stranded nucleic acids (RNA and ssDNA), and neutralization by HIV Vif. The full elucidation of these intermolecular interactions is necessary to understand the mechanism by which these cellular proteins inhibit HIV replication. The role of oligomerization in their activity is an issue of great importance. These are interactions at the nanoscale level, and AFM is the method of choice capable to addressing problems listed above. AFM currently reached a level of development that enables this unique instrument to provide a wealth of important information to the characterization of biomolecular complexes at nanoscale. The topographic analysis of molecular system is the initial area of the AFM application for characterization of the structure and specificity of protein-DNA systems of a broad complexity. In addition to imaging, AFM is capable of direct measurements of molecular interaction within the system and research during this decade led to a dramatic progress in this area of the AFM applications. Our Preliminary Studies have already enabled us to provide the first images of A3G bound to ssDNA substrates. We further anticipate that the use of all modalities of AFM technology will help us detail these complexes, dissect the protein and chemical requirements, and extend key observations to ASF. To help achieve these overarching objectives, we will pursue the following three Specific Aims: Aim 1. Image A3 complexes at nanoscale resolution using AFM (Program Objectives 1 & 2); Aim 2. Measure interactions within A3-ssDNA complexes using force spectroscopy (Program Objectives 1 & 2) and Aim 3. Characterize A3-Vif interactions using AFM imaging and probing modalities (Program Objectives 3). The studies under this project are an integral component of our overall Program Project aiming to provide a comprehensive understanding of A3-AS3 A3-nucleic acid, and A3-Vif interactions. The nanoscale studies will provide a platform for evaluating any future therapeutics that function by leveraging the AS-Vif interaction.

APOBECSG (A3G) 和 APOBECSF (A3F) 参与各种抗病毒蛋白和核酸交易。重要的主要相互作用包括蛋白质间相互作用、结合单链核酸（RNA 和 ssDNA）以及 HIV Vif 的中和。充分阐明这些分子间相互作用对于理解这些细胞蛋白抑制 HIV 复制的机制是必要的。低聚在其活性中的作用是一个非常重要的问题。这些是纳米级水平的相互作用，AFM 是能够解决上述问题的首选方法。 AFM 目前的发展水平使这种独特的仪器能够为纳米级生物分子复合物的表征提供丰富的重要信息。分子系统的形貌分析是 AFM 应用的最初领域，用于表征广泛复杂的蛋白质-DNA 系统的结构和特异性。除了成像之外，AFM 还能够直接测量系统内的分子相互作用，这十年的研究使 AFM 应用这一领域取得了巨大进展。我们的初步研究已经使我们能够提供与 ssDNA 底物结合的 A3G 的第一张图像。我们进一步预计，AFM 技术所有模式的使用将帮助我们详细了解这些复合物，剖析蛋白质和化学要求，并将关键观察扩展到 ASF。为了帮助实现这些总体目标，我们将追求以下三个具体目标：目标 1. 使用 AFM 以纳米级分辨率成像 A3 复合物（计划目标 1 和 2）；目标 2. 使用力谱测量 A3-ssDNA 复合物内的相互作用（计划目标 1 和 2），目标 3. 使用 AFM 成像和探测方式表征 A3-Vif 相互作用（计划目标 3）。该项目下的研究是我们整个计划项目的一个组成部分，旨在全面了解 A3-AS3、A3-核酸和 A3-Vif 相互作用。纳米级研究将为评估任何未来通过利用 AS-Vif 相互作用发挥作用的疗法提供一个平台。