Stabilization of CXCR4 and its complexes for high resolution structural studies

CXCR4 及其复合物的稳定性用于高分辨率结构研究

• 批准号: 7841594
• 负责人: Vadim Cherezov
• 金额: $ 25.73万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-26 至 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7841594
• 关键词: AIDS neuropathy; Acquired Immunodeficiency Syndrome; Address; Adenosine A2A Receptor; Adrenergic Receptor; Agonist; Behavior; Binding; Biological Assay; CCR5 gene; CXCL12 gene; CXCR4 Receptors; CXCR4 gene; Cells; Chemicals; Chemistry; Collaborations; Complex; Crystallization; Data; Dementia; Detergents; Development; Disease; Environment; Environmental Risk Factor; Feedback; G-Protein-Coupled Receptors; Goals; HIV; HIV Envelope Protein gp120; Human; Infection; Joints; Lead; Libraries; Ligand Binding; Ligands; Lipids; Malignant Neoplasms; Measures; Membrane Proteins; Molecular; Phase; Physiological Processes; Process; Production; Property; Proteins; Protocols documentation; Reporting; Research Proposals; Resolution; Roentgen Rays; Role; Sampling; Screening procedure; Specificity; Structural Biologist; Structure; Technology; Time; Toxic effect; United States National Institutes of Health; Variant; Viral; Virus; Work; X ray diffraction analysis; X-Ray Diffraction; analog; chemokine; design; drug development; flexibility; improved; innovation; neuron loss; novel; novel therapeutics; professor; programs; public health relevance; receptor; structural biology; success; three dimensional structure; tool; transmission process

DESCRIPTION (provided by applicant): Experimentally determining the 3-D structure of the G protein-coupled receptor (GPCR), CXCR4 and its complexes, the overall goal of the proposed study, is critical in developing a deeper understanding of its role in HIV viral entry and toxicity; availability of these data is expected to facilitate the development of new therapeutic strategies. Although CCR5 is the primary co-receptor responsible for HIV transmission, emergence of CXCR4-tropic (X4-tropic) viruses later in infection, correlates with a more rapid CD4 decline and a faster progression to AIDS. CXCR4 is also implicated in neuroaids, triggering neuronal cell death and possibly causing onset of dementia. Known conformational flexibility of GPCRs as well as results from preliminary studies have shown that stabilization of the CXCR4 receptor is a significant bottleneck in attempts to crystallize and solve its structures. The two PI's of this research proposal, one a chemist, and the other a crystallographer, propose to develop new chemical tools optimized for structural studies of the HIV co-receptor CXCR4, attempting crystallization and structural determination of the receptors and its complexes. Chemical tools will be designed and optimized for stabilizing, purifying, and detecting the receptor and for measuring its functional behavior. R21 studies will have two aims: (1) Develop tool compounds optimized for structural studies of CXCR4 and (2) Validate new ligands, detergents, and lipids by assessing their impact on protein sample production processes and by producing diffracting crystals of CXCR4. Results from R21 studies will be used to achieve the following in R33 phase of the work: (3) Determine the high-resolution X-ray structure of CXCR4, and (4) Assess the utility of new molecular tools and technologies in stabilizing and crystallizing CXCR4 in complex with gp120. Success of the R21 studies will be achieved by the production of diffracting CXCR4 crystals along with the production of new compound tools that significantly improves receptor sample behavior (e.g. improved stability over time, increased Tm). PUBLIC HEALTH RELEVANCE: Availability of a 3D structure of CXCR4 along with its complexes will lead to developing a deeper understanding of how the HIV virus enters the cell and will help in the design of new therapeutics. CXCR4 is also involved in important physiological process and have been implicated in a number of diseases such as cancer and availability of its structure will likewise have important biomedical implications.

描述（由申请人提供）：通过实验确定 G 蛋白偶联受体 (GPCR)、CXCR4 及其复合物的 3-D 结构，这是拟议研究的总体目标，对于更深入地了解其在 HIV 中的作用至关重要病毒进入和毒性；这些数据的可用性预计将促进新治疗策略的开发。尽管 CCR5 是负责 HIV 传播的主要共同受体，但感染后期出现 CXCR4 向性（X4 向性）病毒，与 CD4 更快下降和更快进展为 AIDS 相关。 CXCR4 还与神经辅助药物有关，引发神经元细胞死亡，并可能导致痴呆症的发作。 GPCR 已知的构象灵活性以及初步研究的结果表明，CXCR4 受体的稳定性是结晶和解析其结构的一个重要瓶颈。该研究计划的两位 PI，一位是化学家，另一位是晶体学家，提议开发新的化学工具，针对 HIV 辅助受体 CXCR4 的结构研究进行优化，尝试对受体及其复合物进行结晶和结构测定。化学工具将被设计和优化以稳定、纯化和检测受体并测量其功能行为。 R21 研究将有两个目标：(1) 开发针对 CXCR4 结构研究优化的工具化合物；(2) 通过评估新配体、去垢剂和脂质对蛋白质样品生产过程的影响以及产生 CXCR4 衍射晶体来验证它们。 R21 研究的结果将用于在 R33 阶段的工作中实现以下目标：(3) 确定 CXCR4 的高分辨率 X 射线结构，以及 (4) 评估新分子工具和技术在稳定和结晶方面的效用CXCR4 与 gp120 形成复合物。 R21 研究的成功将通过生产衍射 CXCR4 晶体以及生产可显着改善受体样品行为（例如随着时间的推移提高稳定性、提高 Tm）的新复合工具来实现。 公共健康相关性：CXCR4 及其复合物的 3D 结构的可用性将导致人们更深入地了解 HIV 病毒如何进入细胞，并将有助于设计新的疗法。 CXCR4还参与重要的生理过程，并与癌症等多种疾病有关，其结构的可用性同样具有重要的生物医学意义。