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）：在实验上确定G蛋白偶联受体（GPCR），CXCR4及其复合物的3-D结构（拟议研究的总体目标）对于对其在HIV病毒进入和毒性中的作用进行更深入的了解至关重要；这些数据的可用性有望促进新的治疗策略的发展。尽管CCR5是负责HIV传播的主要共受体，但在感染后期，CXCR4-热带（X4-纤维化）病毒的出现与CD4的迅速下降和更快的AIDS发展相关。 CXCR4还与神经辅助有关，引发神经元细胞死亡，并可能引起痴呆症发作。 GPCR的已知构象柔韧性以及初步研究的结果表明，CXCR4受体的稳定化是试图结晶和解决其结构的重要瓶颈。该研究建议的两个PI，一个是化学家，另一个是晶体学家，建议开发针对HIV共受体CXCR4的结构研究优化的新化学工具，尝试结晶和受体及其复合物的结构确定。化学工具将被设计和优化，以稳定，净化和检测受体并测量其功能行为。 R21研究将有两个目的：（1）开发针对CXCR4结构研究和（2）通过评估其对蛋白质样品生产过程的影响并通过产生CXCR4衍射晶体来验证新的配体，洗涤剂和脂质的工具化合物。 R21研究的结果将用于在工作的R33阶段中实现以下内容：（3）确定CXCR4的高分辨率X射线结构，（4）（4）评估新分子工具和技术在稳定和与GP120复杂化的CXCR4方面的实用性。 R21研究的成功将通过产生衍射CXCR4晶体以及生产新的化合物工具的生产来实现，从而显着改善受体样本行为（例如，随着时间的推移提高了稳定性，TM提高）。 公共卫生相关性：CXCR4的3D结构及其复合物的可用性将导致对HIV病毒如何进入细胞的更深入了解，并有助于设计新的治疗剂。 CXCR4也参与了重要的生理过程，并且与许多疾病（例如癌症和其结构的可用性）有关，同样将具有重要的生物医学意义。