Massively Parallel Identification of Protein Ligands

蛋白质配体的大规模并行鉴定

• 批准号: 8527800
• 负责人: BENJAMIN F CRAVATT
• 金额: $ 72.6万
• 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: 2009-09-30 至 2015-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8527800
• 关键词: ATP phosphohydrolase; Address; Affinity; Animals; Antibodies; Area; Binding; Biological Assay; Biological Markers; Biological Process; Biology; Biomedical Research; Biopsy; Biotin; Cells; Chemicals; Communities; Complex; Diagnostic; Disease; Drug Formulations; Enzymes; Epitopes; Fluorescence; Goals; Gold; Health; Human; Label; Laboratories; Libraries; Ligands; Link; Mass Spectrum Analysis; Medicine; Methods; Monitor; Monoclonal Antibodies; N-substituted Glycines; Nucleic Acids; Organic Chemistry; Peptoids; Pharmaceutical Preparations; Phosphotransferases; Process; Protein Binding; Proteins; Proteome; Proteomics; Reaction; Reagent; Research; Research Project Grants; Role; Ruthenium; Sampling; Serine Hydrolase; Serum; Singlet Oxygen; Sorting - Cell Movement; Source; Specificity; Techniques; Technology; Tissue Extracts; Tissues; United States National Institutes of Health; Visible Radiation; activity-based protein profiling; analog; appendage; aptamer; base; combinatorial; frontier; inhibitor/antagonist; inorganic phosphate; interest; member; novel; oxidation; screening; skills; tool

DESCRIPTION (provided by applicant): This project aims to develop a revolutionary screening platform that will allow for the rapid isolation of hundreds of high affinity and specificity synthetic ligands for proteins in a highly parallel fashion. The central feature of the proposal is to label dozens to hundreds of mechanistically related proteins simultaneously using reagents developed for Activity-Based Protein Profiling (ABPP). This mixture of labeled proteins will then be screened en masse against a several million member combinatorial library of peptoids displayed on beads. Peptoid-displaying beads that retain labeled protein will be collected by fluorescence-activated flow sorting. Mass spectrometry-based techniques will then be employed to identify the protein captured on the bead as well as the sequence of the peptoid. These peptoids can be employed as capture agents in the creation of protein-detecting microarrays. They will also be modified for use as high potency, photo-triggered pharmacological inhibitors of their target proteins. This will involve appendage of a Ru(II)-containing complex to the peptoid. When irradiated with visible light, the ruthenium complex generates singlet oxygen, which is capable of destroying proteins in the immediate vicinity. These novel reagents will be used to probe the roles of the target proteins in biological assays. If successful, this effort will revolutionize the discovery of specific protein ligands, a goal identified by the NIH as a strategic priority. Because the ligands will be peptoids, which can be easily synthesized in large quantities even by laboratories lacking specialized organic chemistry skills, these ligands will be far more widely accessible to the research community than would be the case for most other classes of protein binding-molecules. Therefore, we believe that this project is transformative in its potential scope and impact. PUBLIC HEALTH RELEVANCE: The identification of large numbers of protein ligands has been identified by the NIH as a high priority for biomedical research. Such ligands could be employed as reagents to construct tools for the discovery of diagnostically useful disease biomarkers. The synthetic compounds that we plan to identify could also serve as drug leads for a variety of therapeutically interesting targets.

描述（由申请人提供）：该项目旨在开发一个革命性的筛选平台，该平台将允许以高度平行的方式快速隔离数百种高亲和力和特异性合成配体的蛋白质。该提案的主要特征是使用用于基于活性的蛋白质谱（ABPP）的试剂同时将数十个机械性相关的蛋白质标记为数百种机械性相关的蛋白质。然后将将标记蛋白的这种混合物大批筛选，以在珠子上显示的几百万个肽库组合库。保留标记蛋白的肽滴定珠将通过荧光激活的流动分类收集。然后将采用基于质谱的技术来识别珠子上捕获的蛋白质以及肽的序列。这些肽可以用作蛋白质检测微阵列的捕获剂。它们还将被修饰，以作为其靶蛋白的高效力，光触发的药理抑制剂。这将涉及将RU（II）含有肽的复合物附加到肽中。当用可见光照射时，芳族复合物会产生单线氧，该氧气能够破坏附近的蛋白质。这些新型试剂将用于探测靶蛋白在生物测定中的作用。如果成功的话，这项工作将彻底改变发现特定蛋白质配体的发现，这是NIH确定为战略优先事项的目标。由于配体将是肽，即使缺乏专门的有机化学技能的实验室也可以很容易地合成，因此研究界将比大多数其他类别的蛋白质结合 - 分子更广泛地使用这些配体。因此，我们认为该项目的潜在范围和影响是变革性的。 公共卫生相关性：NIH已将大量蛋白质配体的鉴定为生物医学研究的重点。这种配体可以用作试剂来构建用于发现诊断有用的疾病生物标志物的工具。我们计划鉴定的合成化合物也可以作为各种治疗靶标的药物铅。

项目成果

A liquid array platform for the multiplexed analysis of synthetic molecule-protein interactions.

• DOI: 10.1021/cb400806r
• 发表时间: 2014-02-21
• 期刊: ACS CHEMICAL BIOLOGY
• 影响因子: 4
• 作者: Doran, Todd M.;Kodadek, Thomas
• 通讯作者: Kodadek, Thomas

Solid phase synthesis of 1,3,4-oxadiazin-5 (6R)-one and 1,3,4-oxadiazol-2-one scaffolds from acyl hydrazides.

• DOI: 10.1039/c4ob01883d
• 发表时间: 2015-01-07
• 期刊: Organic & biomolecular chemistry
• 影响因子: 3.2
• 作者: Sarma BK;Liu X;Wu H;Gao Y;Kodadek T
• 通讯作者: Kodadek T

Rapid development of a potent photo-triggered inhibitor of the serine hydrolase RBBP9.

• DOI: 10.1002/cbic.201200445
• 发表时间: 2012-09-24
• 期刊: CHEMBIOCHEM
• 影响因子: 3.2
• 作者: Liu, Xiaodan;Dix, Melissa;Speers, Anna E.;Bachovchin, Daniel A.;Zuhl, Andrea M.;Cravatt, Benjamin F.;Kodadek, Thomas J.
• 通讯作者: Kodadek, Thomas J.

Incorporation of heterocycles into the backbone of peptoids to generate diverse peptoid-inspired one bead one compound libraries.

• DOI: 10.1021/co200195t
• 发表时间: 2012-03-12
• 期刊: ACS combinatorial science
• 作者: Aditya A;Kodadek T
• 通讯作者: Kodadek T