Systematic stabilization of specific protein-protein interactions

特定蛋白质-蛋白质相互作用的系统稳定

基本信息

批准号：
10703416
负责人：
Michelle Arkin
金额：
$ 31.67万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-09-15 至 2026-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10703416
关键词：
Adaptor Signaling Protein Address Binding Binding Proteins Binding Sites Biological Biological Models Biological Process Biology Cell Cycle Cell Cycle Progression Cell Cycle Regulation Pathway Cells Cellular Assay Chemicals Client Complex Development Disease Disulfides Estrogen Receptor alpha Estrogen Receptors FOXO1A gene Genetic Transcription Glues Goals Homodimerization Human Individual Knowledge Learning Length Link Malignant Neoplasms Methodology Methods Modification Molecular Molecular Conformation Multiprotein Complexes Natural Products Nature Nerve Degeneration Outcome Peptides Pharmaceutical Preparations Phosphopeptides Phosphorylation Phosphotransferases Physiological Processes Play Positioning Attribute Protac Protein Isoforms Proteins Proteome Role Scaffolding Protein Serine Signal Transduction Signal Transduction Pathway Site Specificity Structure Surface System Technology Testing Therapeutic Threonine Transcriptional Regulation Translating design drug discovery drug-like compound enzyme activity estrogen-related receptor flexibility fusicoccin immune modulating agents inhibitor insight iterative design non-Native novel novel therapeutics prospective protein complex protein protein interaction proteostasis raf Kinases rare genetic disorder scaffold screening small molecule synthetic protein tool transcription factor ubiquitin ligase

项目摘要

Abstract Protein-protein interactions (PPIs) are ubiquitous in biology, and their dysregulation is closely associated with diseases, from cancer to neurodegeneration to rare genetic disorders. PPIs often form complex networks that include highly interacting ‘hub’ proteins. Methods to modulate single interactions would provide great insight into the functions of these hubs. Small-molecule probes and drug leads have focused on blocking PPIs; however, stabilizing PPIs could be just as important for drug discovery and could provide greater selectivity for chemical biology. However, there are few systematic methodologies to discover PPI stabilizers prospectively. This proposal focuses on the systematic discovery of selective small-molecule PPI stabilizers, using the hub protein 14-3-3 as a model system. 14-3-3s are seven highly homologous adaptor proteins that bind to serine and threonine sites on client proteins to alter their function and fate. Hundreds of proteins in signal transduction pathways, cell-cycle regulation, transcription regulation, and protein homeostasis are clients of 14-3-3. Given the importance of protein phosphorylation and the ubiquity of 14-3-3 as an effector of phosphorylation, it is surprisingly underappreciated. We propose that developing a tool kit of selective, cell-active stabilizers of native 14-3-3/client PPIs will stimulate biological study and may lead to new drugs. Based on the structural diversity of clients, we hypothesize that we can develop client-selective PPI stabilizers that bind to the composite 14-3- 3/client interface. These selective stabilizers should amplify the native biology of the 14-3-3/client complex. We will provide proof-of-concept for this approach through three aims: Aim 1. Screen for selective stabilizers of 14-3-3/phosphopeptide clients. We have previously discovered disulfide-bound fragments that stabilize 14-3- 3/phosphopeptide complexes. We will now screen six, structurally and biologically diverse 14-3-3/client complexes, using a native C38 residue found only on the 14-3-3 isoform. We hypothesize that client sequences with more open or flexible structures near the C38 will yield higher quality hits. Aim 2. Optimize 14-3-3/client stabilizers for cell-based activity. We have demonstrated the ability to convert disulfides to cell-active electrophilic warheads and to tune the selectivity of 14-3-3/client stabilizers. We will optimize C38-bound fragments with (or without) an electrophile with the goal of achieving target-selective PPI stabilization in cells for the 14-3-3 clients CRAF kinase, estrogen receptor  (ER), and the transcription factor FOXO1. Aim 3. Design PROTAC-based degraders of 14-3-3 clients. PROTACs are bifunctional molecules that induce proximity between a ubiquitin ligase and a target, leading to the target’s degradation. We will expand the targets accessible to PROTAC technology by using 14-3-3 as a scaffolding protein to link intrinsically disordered proteins (IDPs) to the ubiquitin ligase. We will first develop the technology for ER, where PROTACs are known, then translate our learning to IDPs, including FOXO1. Successful completion of these aims will provide approaches that are broadly applicable to the 14-3-3 network and are extendable to other native and nonnative PPIs.

抽象的蛋白质-蛋白质相互作用（PPI）在生物学中普遍存在，其失调与从癌症到神经退行性疾病再到罕见的遗传性疾病，质子泵抑制剂（PPI）通常形成复杂的网络。包括高度相互作用的“中心”蛋白质，调节单一相互作用的方法将为我们提供深入的了解。然而，这些中枢的功能主要是阻断 PPI；稳定 PPI 对于药物发现同样重要，并且可以为化学药物提供更大的选择性然而，很少有系统的方法来前瞻性地发现 PPI 稳定剂。该提案的重点是使用 hub 蛋白系统地发现选择性小分子 PPI 稳定剂 14-3-3 作为模型系统，是与丝氨酸和 14-3-3 结合的七个高度同源的接头蛋白。客户蛋白质上的苏氨酸位点可改变其信号转导中的数百种蛋白质的功能和命运。通路、细胞周期调控、转录调控和蛋白质稳态是 14-3-3 的客户。蛋白质磷酸化的重要性以及 14-3-3 作为磷酸化效应子的普遍存在，令人惊讶的是，我们建议开发一种选择性的、细胞活性的天然稳定剂工具包。 14-3-3/客户 PPI 将刺激生物学研究，并可能产生基于结构多样性的新药物。客户，我们认为我们可以开发客户选择性的 PPI 稳定剂，与复合材料 14-3- 结合 3/客户界面。这些选择性稳定剂应该增强 14-3-3/客户复合物的天然生物学特性。将通过三个目标为这种方法提供概念验证：目标 1. 筛选选择性稳定剂 14-3-3/磷酸肽客户之前发现了可稳定 14-3- 的二硫键结合片段。 3/磷酸肽复合物现在将筛选六种结构和生物多样性的14-3-3/客户。复合物，使用仅在 14-3-3 同工型上发现的天然 C38 残基，我们与该客户序列进行了斗争。在 C38 附近采用更开放或更灵活的结构将产生更高质量的命中。目标 2. 优化 14-3-3/客户端。我们已经证明了将二硫化物转化为细胞活性的稳定剂。我们将优化 C38 结合的亲电子弹头并调整 14-3-3/客户稳定剂的选择性。具有（或不具有）亲电子试剂的片段，目的是在细胞中实现目标选择性 PPI 稳定 14-3-3 客户端 CRAF 激酶、雌激素受体 α (ERα) 和转录因子 FOXO1。目标 3。基于 PROTAC 的 14-3-3 客户端降解剂是诱导接近的双功能分子。泛素连接酶和靶标之间的连接，导致靶标降解。我们将扩大可访问的靶标。 PROTAC 技术，使用 14-3-3 作为支架蛋白，将本质无序蛋白 (IDP) 连接到我们将首先开发 ERα 技术，其中 PROTAC 是已知的，然后转化我们的技术。向国内流离失所者学习，包括 FOXO1，成功完成这些目标将为您提供广泛的方法。适用于 14-3-3 网络，并可扩展到其他本地和非本地 PPI。

项目成果

期刊论文数量（3）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Reversible Dual-Covalent Molecular Locking of the 14-3-3/ERRγ Protein-Protein Interaction as a Molecular Glue Drug Discovery Approach.

DOI：
10.1021/jacs.2c12781
发表时间：
2023-03-29
期刊：
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY
影响因子：
15
作者：
Somsen, Bente A.;Schellekens, Rick J. C.;Verhoef, Carlo J. A.;Arkin, Michelle R.;Ottmann, Christian;Cossar, Peter J.;Brunsveld, Luc
通讯作者：
Brunsveld, Luc

Structure-Based Optimization of Covalent, Small-Molecule Stabilizers of the 14-3-3σ/ERα Protein-Protein Interaction from Nonselective Fragments.

DOI：
10.1021/jacs.3c05161
发表时间：
2023-09-20
期刊：
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY
影响因子：
15
作者：
Konstantinidou, Markella;Visser, Emira J.;Vandenboorn, Edmee;Chen, Sheng;Jaishankar, Priyadarshini;Overmans, Maurits;Dutta, Shubhankar;Neitz, R. Jeffrey;Renslo, Adam R.;Ottmann, Christian;Brunsveld, Luc;Arkin, Michelle R.
通讯作者：
Arkin, Michelle R.

From Tethered to Freestanding Stabilizers of 14-3-3 Protein-Protein Interactions through Fragment Linking.

通过片段连接从 14-3-3 蛋白质-蛋白质相互作用的束缚稳定剂到独立稳定剂。

DOI：
10.1002/anie.202308004
发表时间：
2023
期刊：
Angewandte Chemie (International ed. in English)
影响因子：
0
作者：
Visser,EmiraJ;Jaishankar,Priyadarshini;Sijbesma,Eline;Pennings,MarloesAM;Vandenboorn,EdmeeMF;Guillory,Xavier;Neitz,RJeffrey;Morrow,John;Dutta,Shubhankar;Renslo,AdamR;Brunsveld,Luc;Arkin,MichelleR;Ottmann,Christian
通讯作者：
Ottmann,Christian