Protein technologies

蛋白质技术

• 批准号: 7667729
• 负责人: IRWIN M CHAIKEN
• 金额: $ 19.51万
• 依托单位: DREXEL UNIVERSITY
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/7667729
• 关键词: Antibodies; Antiviral Agents; Area; Binding; Biological Assay; CD4 Antigens; CXCR4 gene; Calorimetry; Cell Line; Cells; Chemokine (C-C Motif) Receptor 5; Computer Simulation; Crystallography; Data; Development; Event; Fc Receptor; Goals; HIV Envelope Protein gp120; HIV-1; Infection; Investigation; Kinetics; Label; Ligand Binding; Ligands; Methodology; Methods; Molecular; Numbers; Peptides; Principal Investigator; Process; Program Research Project Grants; Property; Protein Binding; Proteins; Quantitative Evaluations; Range; Reagent; Recombinant Proteins; Resources; Screening procedure; Services; Structure; Surface Plasmon Resonance; System; Technology; Testing; Thermodynamics; Viral; Virus; Work; base; chemokine receptor; design; driving force; env Gene Products; inhibitor/antagonist; interdisciplinary approach; member; method development; monomer; mutant; new technology; novel; programs; protein expression; receptor; size; small molecule

Principal Investigator/Program Director (Last, First, Middle): Chaiken, IfWJn M DESCRIPTION: The main objective of Core B is to provide protein technology services, including protein reagents, ligand binding analyses and methods development, to support investigation of structure-based antagonism of human immunodeficiency virus -1 (HIV-1) entry. HIV-1 infection is initiated by entry of the virus into host cells. Interaction of HIV-1 gp120 envelope protein with host cell receptor, CD4, followed by binding to a co-receptor, most commonly the chemokine receptors CCR5 or CXCR4, are crucial steps in the process of viral entry. The overall goal of this Program Project is to identify agents that will antagonize these protein-binding events. Candidate inhibitors of gp120 interactions will be generated by various members of the Program Project team, in particular, Project 1, Structure analysis of HIV-1 entry inhibition, Project 2, Peptide-inspired competitive and allosteric inhibitors of HIV-1 entry and Project 3, Discovery and synthesis of small-molecule CD4-gp120 antagonists. In order to analyze the efficacy and mechanisms of action of these candidate inhibitors, protein reagents will be required by the above projects, in coordination with Project 4, Assembly and inhibition thermodynamics, and Project 5, Structure-based antagonism of HIV-1 envelope function in cell entry. Furthermore, SPR-based screening of these inhibitor candidates with gp120s from multiple subtypes, both in direct binding and functional assays, will help prioritize thermodynamic investigations, while also aiding the molecular interpretation of biological assays. The data generated through the screening assays also will help test the hypotheses of inhibitor interaction mechanisms developed in Core A, Computational modeling. The specific aims of Core B are to: (1) Produce, purify and validate protein reagents required by Program Project members, including envelope monomers and trimers from diverse clades of HIV-1, soluble constructs of HIV-1 receptors and antibodies for both purification and characterization studies. (2) Screen kinetic interaction properties of gp120 antagonist candidates for direct interaction and effector activity profiles, which will contribute information that will be invaluable in the compound prioritization process. (3) Develop, validate and implement new technologies for recombinant protein expression, purification and binding assays to serve evolving needs of the Program Project. Transfer new technologies to P01 projects to further enable mechanistic and antagonist design investigations. Overall, this Core will provide Project Team members with a central resource that will both produce and supply high quality protein reagents, in addition to screening candidate antiviral molecules and developing novel assay systems for the investigation of these compounds. The work of Core B will adapt its activities in these areas to the evolving needs of the Projects in order to support investigations of HIV-1 entry antagonism and structure- based antagonist design.

首席研究员/计划主任（最后，第一，中间）：chaiken，ifwjn m 描述： 核B的主要目的是提供蛋白质技术服务，包括蛋白质试剂，配体结合 分析和方法开发，以支持对人类基于结构的拮抗作用的研究 免疫缺陷病毒-1（HIV -1）进入。 HIV-1感染是通过将病毒进入宿主细胞引发的。相互作用 带有宿主细胞受体CD4的HIV-1 GP120包膜蛋白，然后与共受体结合，大多数 通常，趋化因子受体CCR5或CXCR4是病毒进入过程中的关键步骤。总体 该计划项目的目标是确定将拮抗这些蛋白质结合事件的代理。候选人 GP120互动的抑制剂将由计划项目团队的各个成员产生 项目1，HIV-1进入抑制的结构分析，项目2，肽启发的竞争性和变构性 HIV-1进入和项目3的抑制剂，小分子CD4-GP120拮抗剂的发现和合成。在 为了分析这些候选抑制剂的作用疗效和机制，蛋白质试剂将是 上述项目要求，与项目4，组装和抑制热力学以及 项目5，基于结构的HIV-1信封功能在细胞进入中的拮抗作用。此外，基于SPR 通过直接结合和功能性筛选使用多种亚型的GP120候选者筛选 测定，将有助于优先进行热力学研究，同时也有助于分子解释 生物测定。通过筛选测定产生的数据也将有助于测试抑制剂的假设 在核心A，计算建模中开发的交互作用机制。 核心B的具体目的是：（1）生产，净化和验证程序项目所需的蛋白质试剂 HIV-1各种进化枝的信封单体和三聚体，包括HIV-1的可溶性结构 用于纯化和表征研究的受体和抗体。 （2）屏幕动力学相互作用属性 GP120的拮抗剂候选者的直接相互作用和效应活动曲线，这将贡献信息 在复合优先级过程中，这将是无价的。 （3）开发，验证和实施新的 重组蛋白表达，纯化和结合测定的技术，以满足不断发展的需求 计划项目。将新技术转移到P01项目，以进一步启用机械和对手设计 调查。 总体而言，该核心将为项目团队成员提供既有生产又提供供应的中心资源 高质量的蛋白质试剂除了筛选候选抗病毒分子并开发新的测定法 研究这些化合物的系统。核心B的工作将调整其在这些领域的活动 项目的不断发展的需求是为了支持对HIV-1进入对抗和结构的调查 - 基于拮抗剂设计。