MINIPROTEIN MIMETICS

小蛋白模拟物

• 批准号: 6658421
• 负责人: IRWIN M CHAIKEN
• 金额: $ 15.72万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-09-10 至 2003-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6658421
• 关键词: CD4 molecule; HIV envelope protein gp120; HIV infections; T lymphocyte; X ray crystallography; antiAIDS agent; binding sites; biomimetics; cytokine receptors; drug design /synthesis /production; human immunodeficiency virus 1; liposomes; nuclear magnetic resonance spectroscopy; peptide chemical synthesis; protein engineering; protein protein interaction; protein structure; recombinant proteins; site directed mutagenesis; synthetic protein

DESCRIPTION (provided by applicant): The central objective of this Project is to use miniprotein mimetics together with the structure of the gp120KD4 complex to design antagonists of the interaction of HIV-1 with human cells. T-cell docking and entry by HIV-1, a major route of cell infection in AIDS, is driven by specific recognition of T-cell surface protein CD4 and chemokine coreceptor by HIV envelope protein gp120. The crystallographic structure of CD4 is known, as is its complex with gp120 and the coreceptor surrogate antibody 17b. The structural nature and interrelationship of gp 120 binding sites for CD4 and coreceptor have been revealed. The advancing high-resolution structural understanding of the protein participants in virus-cell recognition, together with the advancing technology of mimetics design, now make it possible to use miniprotein engineering coordinately with biophysical and biologicalanalysis to obtain an advanced mechanistic understanding of the structural basis of the CD4-gp120interaction process. This mechanistic understanding will help guide the design of new antagonists for HIV infection. The specific aims are: (1) Construct novel miniprotein mimetics of CD4 using synthetic and complementary phage randomization methodologies and optimize their affinities for HIV- 1 envelope; (2) Identify key structural elements in miniprotein mimetics of CD4 that control activation of the coreceptor binding site of HIV-1 Env, and form miniprotein variants which favor Env binding with diminished coreceptor site activation; (3) Determine structural and interaction properties of miniprotein mimetics with trimeric vs monomeric forms of HIV- 1 envelopes, including ectodomain and membrane-associated forms, and refine the topological map of key structural elements needed for HIV-1-cell attachment and attachment antagonists. Overall, this project will yield an advanced definition of key structural elements at the interface of the CD4-gp 120 complex that trigger increased HIV- 1 envelope affinity for co-receptor and consequent cell infection; improved understanding of the CD4-viral envelope-coreceptor molecular machine that drives HIV- 1 attachment and infection of host cells; and molecular leads for new antagonist candidates for antiviral therapy. The miniprotein mimetics strategies derived will be useful to determine the mechanism of, and design antagonists for, other protein-protein interactions, such as the gp41 -gp 120 and chemokine receptor.

描述（由申请人提供）：该项目的核心目的是使用微蛋白 Mimetics与GP120KD4复合物的结构一起设计HIV-1与人类细胞相互作用的拮抗剂。 HIV-1是AIDS中的主要细胞感染途径HIV-1的T细胞对接和进入，这是由对T细胞表面蛋白CD4的特异性识别和HIV包膜蛋白GP120的T细胞表面蛋白CD4和趋化因子共肽的驱动。 CD4的晶体结构是已知的，它与GP120和croecector替代抗体17b的复合物也是如此。已经揭示了CD4和CORECEPTOR的GP 120结合位点的结构性质和相互关系。对病毒细胞识别中蛋白质参与者的高分辨率结构理解，以及Mimetics Design的前进技术，现在可以协调使用微蛋白工程与生物物理和生物学分析，以获得对CD4-GP120 Interaction过程的结构基础的高级机械理解。这种机械理解将有助于指导新对手的设计 艾滋病毒感染。具体目的是：（1）使用合成和互补的噬菌体随机方法来构建CD4的新型微型蛋白模拟物，并优化其对HIV-1包膜的亲和力； （2）在CD4的微蛋白模拟物中确定关键的结构元素，以控制HIV-1 Env的共感受器结合位点的激活，并形成微型蛋白质变体，这些变体有利于env eNV结合降低的corecector seret活化； （3）确定与三聚体与单体形式的HIV-1个信封的结构和相互作用特性，包括外生域和膜相关形式，并完善HIV-1细胞附着和附着拮抗剂所需的关键结构元素的拓扑图。总体而言，该项目将在CD4-GP 120复合物界面的关键结构元素中产生高级定义，从而触发了HIV-1包络线对共受体和随之而来的细胞感染的增强；对CD4-病毒包膜可感染的分子机器的理解得到了改善，该机器驱动HIV-1的附着和宿主细胞感染；新的抗病毒治疗替代疗法的分子铅。得出的小巧蛋白模拟策略将有助于确定其他蛋白质蛋白相互作用（例如GP41 -GP 120和趋化因子受体）的机理和设计拮抗剂的机理。