A Lexicon of Stapled Peptide Helices Engineered to Capture the Protein Interactom

旨在捕获蛋白质相互作用的钉合肽螺旋词典

• 批准号: 7937806
• 负责人: Loren David Walensky
• 金额: $ 43.27万
• 依托单位: DANA-FARBER CANCER INST
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-30 至 2014-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7937806
• 关键词: Affinity; Binding; Binding Sites; Biological; Biology; Cataloging; Catalogs; Cells; Chemicals; Chemistry; Clinical; Disease; Disease Pathway; Drug Design; Drug Interactions; Engineering; Goals; Health; Human; Hydrocarbons; Immobilization; Implant; Link; Mediating; Medicine; Molecular; Molecular Structure; Nature; Peptides; Proteins; Proteomics; Recreation; Research Infrastructure; Seminal; Shapes; Site; Specificity; Technology; Therapeutic Intervention; Tooth structure; Translations; base; grasp; high throughput technology; in vivo; intercalation; interdisciplinary approach; novel; peptide structure; protein complex; public health relevance; research study

DESCRIPTION (provided by applicant): Whether fleeting or stable, normal or aberrant, protein interactions and their sites of contact form the basis for discovery of biological pathways, disease mechanisms, and opportunities for therapeutic intervention. The goal of this proposal is to intertwine chemistry, biology, and medicine to create a transformative high- throughput technology that precisely identifies protein targets and their explicit sites of interaction. Like the teeth of a key that perfectly fit into a lock, complementary protein shape is critical to the execution of biological interactions. The molecular handshakes of proteins rely on their discrete substructures and these contact points are typically embedded within a complex protein that provides the infrastructure to maintain the essential bioactive fold. Ideally, these evolutionarily honed substructures could be used to capture and thereby catalogue their protein targets; however, out of context from the whole protein, bioactive subdomains often unfold, resulting in loss of biological shape, potency, and specificity. To reclaim the enormous capacity of structured peptides to selectively bind and capture their protein targets, we will first restore their bioactive shape and then chemically derivatize them for both non-covalent and covalent capture. In this proposal, we focus on the peptide 1-helix, arguably the most ubiquitous and versatile biological shape harnessed by the cell. We will apply our robust "hydrocarbon stapling" technology to synthesize a diversity of bioactive 1-helices and then chemically install new immobilization and intercalating functionalities to expand our grasp of the interactome by trapping the full-range of stable to transient protein interactors. Our covalent capture chemistry and proteomic analyses afford a two-for-one advantage: identification of protein targets and their sites of interaction. Since protein interaction sites are the topographic templates for drug design, we believe that the binding site identification feature of our approach will provide a critical link between interactome discovery and clinical translation. To accomplish our goals, we will take a step-wise approach: (1) structural stabilization, (2) directional affinity capture, (3) covalent capture, and (4) binding site identification. Each step will be adapted for high-throughput and validated using proof-of-concept biological experiments. Once identified and catalogued, protein interactions must be validated biologically. A seminal feature of our approach is that the very 1-helices we use to capture the protein interactome can be used to validate and drug the interactions in cellular and in vivo studies. Thus, we believe that engineering stapled peptides for protein capture will create a powerful and versatile approach to elucidating the interactome, and massively expand the potential for discovery of novel interactions and how they impact health and disease. PUBLIC HEALTH RELEVANCE: Protein interactions mediate innumerable cellular activities in health and disease; our goal is to create a transformative high-throughput technology that rapidly and precisely identifies protein targets and their explicit sites of interaction. The novelty of our multidisciplinary approach begins with the chemical recreation of protein substructures that mediate protein interaction, transforming Nature's evolutionarily-honed binding motifs into a discovery toolbox; next, we chemically implant in these bioactive structures molecular functionalities for immobilization and irreversible protein intercalation. By operating at the interface of chemistry, biology, and medicine, we aim to develop and deploy a technology that surmounts the formidable challenge of identifying, distinguishing, and drugging the broad array of human protein targets.

描述（由申请人提供）：无论是短暂的还是稳定，正常或异常，蛋白质相互作用及其接触部位构成了发现生物途径，疾病机制和治疗干预的机会的基础。该建议的目的是交织化学，生物学和医学，以创建一种变革性的高吞吐技术，以精确识别蛋白质靶标及其显式相互作用。就像完全适合锁定的钥匙的牙齿一样，互补的蛋白质形状对于执行生物学相互作用至关重要。蛋白质的分子握手依赖于它们的离散子结构，这些接触点通常嵌入复杂的蛋白质中，该蛋白质提供了基础结构以维持必需的生物活性折叠。理想情况下，这些进化磨练的子结构可用于捕获并从而对其蛋白质靶标进行分类。然而，在整个蛋白质的上下文中，生物活性亚域通常会展开，从而导致生物形状，效力和特异性的丧失。为了收回结构化肽选择性结合和捕获其蛋白质靶标的巨大能力，我们将首先恢复其生物活性形状，然后化学衍生它们，以既非共价和共价捕获。在此提案中，我们专注于肽1-螺旋，可以说是细胞利用的最普遍和多功能的生物形状。我们将应用强大的“碳氢化合物钉”技术来综合多样的生物活性1螺旋，然后化学安装新的固定化和插入功能，从而扩大我们对相互作用组的掌握，通过捕获稳定的稳定范围为瞬时蛋白质相互作用者。我们的共价捕获化学和蛋白质组学分析具有两对一优势：识别蛋白质靶标及其相互作用位点。由于蛋白质相互作用位点是药物设计的地形模板，因此我们认为我们方法的结合位点识别特征将在相互作用的发现与临床翻译之间提供关键的联系。为了实现我们的目标，我们将采用逐步的方法：（1）结构稳定，（2）方向亲和力捕获，（3）共价捕获和（4）结合位点识别。每个步骤都将适应高通量，并使用概念验证生物学实验进行验证。一旦确定并分类，蛋白质相互作用就必须在生物学上进行验证。我们方法的开创性特征是，我们用来捕获蛋白质相互作用组的1个螺旋可用于验证和吸毒细胞和体内研究中的相互作用。因此，我们认为，用于蛋白质捕获的工程钉肽将创建一种强大而多才多艺的方法来阐明相互作用，并大大扩展了发现新型相互作用以及它们如何影响健康和疾病的潜力。 公共卫生相关性：蛋白质相互作用介导了健康和疾病中无数的细胞活动；我们的目标是创建一种变革性的高通量技术，该技术可以快速并精确地识别蛋白质靶标及其显式相互作用。我们多学科方法的新颖性始于介导蛋白质相互作用的蛋白质亚结构的化学娱乐，将自然的进化结合基序转化为发现工具箱。接下来，我们在这些生物活性结构中化学植入分子功能，以固定和不可逆的蛋白质插入。通过在化学，生物学和医学的界面上运行，我们旨在开发和部署一项技术，以探索识别，区分和吸毒的巨大挑战。