Chemical Modulation of the Siah-1 Pathway

Siah-1 通路的化学调节

基本信息

批准号：
7694153
负责人：
JOHN C REED
金额：
$ 2.5万
依托单位：
SANFORD BURNHAM PREBYS MEDICAL DISCOVERY INSTITUTE
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-04-15 至 2010-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7694153
关键词：
Apoptosis Binding Binding Sites Biological Biological Assay Biology Breast Cancer Cell Cancer cell line Cell Cycle Arrest Cell Cycle Regulation Cell Line Cell division Cell physiology Cells Chemicals Chemistry Complex Consensus Cytokinesis DNA Damage Data Degradation Pathway Development Engineering Enzymes Event Excision Family Fibroblasts Fluorescence Polarization Genes Goals Human Hypoxia Laboratories Libraries Ligands Link Luciferases Malignant Neoplasms Mammalian Cell Mediating Mediator of activation protein Messenger RNA Methodology Methods Mitotic Mixed Function Oxygenases Mus Mutation N-terminal Pathway interactions Peptides Plasmids Play Polyubiquitin Post-Translational Protein Processing Process Production Proline Protein Family Proteins Proteolysis Provider Recovery Reporter Genes Reporting Research Role Screening procedure Serum Siah-1 protein Signal Pathway Signal Transduction Structure-Activity Relationship TCF Transcription Factor TP53 gene Tertiary Protein Structure Transcription Coactivator Tubulin Tumor Suppression Tumor Suppressor Proteins Ubiquitin Ubiquitin-Conjugating Enzymes Ubiquitination United States National Institutes of Health Vav guanine-nucleotide exchange factor Work base cell growth cell transformation cellular engineering cheminformatics high throughput screening human NCOR1 protein malignant stomach neoplasm member metabotropic glutamate receptor 4 multicatalytic endopeptidase complex overexpression pancreatic neoplasm promoter protein degradation protein protein interaction public health relevance response small molecule libraries therapy resistant tool tumor tumor growth ubiquitin ligase ubiquitin-protein ligase

项目摘要

DESCRIPTION (provided by applicant): Proteasomal degradation typically requires post-translational modification of target proteins with K48-linked polyubiquitin chains. This process of protein proteolysis plays a key role in normal cellular function. The E3 ubiquitin ligase, Siah-1, facilitates the transfer of ubiquitin to its substrate proteins destined for degradation by way of its RING domain. Siah-1 is a member of a family of highly conserved RING domain proteins, which regulate a variety of cellular functions, including cell cycle arrest, tumor suppression, and apoptosis through the -catenin degradation pathway. Siah-1 has also been identified as a p53-inducible gene, functionally linking it to an important tumor suppressor. Chemical modulators of the Siah-1 pathway would provide powerful research tools for elucidating the roles of this signaling pathway in cancer development and progression. In this proposal, we describe the development of a High Throughput Screening (HTS) assay based upon fluorescence polarization, and utilizing a peptide ligand of Siah-family proteins with an attached flurochrome. This fluorescence polarization assay (FPA) forms the basis for a high-throughput competitive displacement assay that we have optimized for chemical library screening. We propose to screen the NIH compound library using this HTS assay. Additional downstream assays provided by the assay provider will be performed for deconvoluting hits. We expect to obtain candidate compounds for Structure Activity Relationship (SAR) studies to be performed for a prototypical RING- containing protein, Siah-1. Together, these efforts will result in validated chemical probes for studying the biology of Siah-family E3 ligases in a variety of biological settings. PUBLIC HEALTH RELEVANCE: Our goal is to identify chemicals capable of modulating the Siah-1 proteasomal degradation pathway. Through its activity as an ubiquitin ligase, this signaling pathway helps to regulate a variety of cellular events, including cell cycle arrests, tumor suppression, and apoptosis. Chemical modulators of the Siah-1 pathway will provide powerful research tools for studying pathways involved in human cancer development, progression, and resistance to therapy.

描述（由申请人提供）：蛋白酶体降解通常需要用K48连接的多泛素链对靶蛋白进行翻译后修饰。这种蛋白质蛋白水解的过程在正常细胞功能中起关键作用。 E3泛素连接酶SIAH-1促进了泛素通过其环域转移到其底物蛋白上的降解。 SIAH -1是高度保守的环域蛋白家族的成员，该家族通过-Catenin降解途径来调节各种细胞功能，包括细胞周期停滞，肿瘤抑制和凋亡。 SIAH-1也已被鉴定为p53诱导的基因，在功能上将其与重要的肿瘤抑制剂联系起来。 SIAH-1途径的化学调节剂将为阐明该信号通路在癌症发育和进展中的作用提供强大的研究工具。在此提案中，我们描述了基于荧光偏振的高吞吐量筛选（HTS）测定的发展，并利用带有附着的氟色素的SIAH-家庭蛋白质的肽配体。该荧光偏振分析（FPA）构成了我们为化学库筛选优化的高通量竞争位移测定法的基础。我们建议使用此HTS测定法筛选NIH复合库。分析提供商提供的其他下游测定法将用于击球效果。我们期望为含有原型环的蛋白质SIAH-1进行结构活性关系（SAR）研究的候选化合物（SAR）研究。这些努力将共同研究在各种生物环境中研究SIAH家庭E3连接酶的生物学的经过验证的化学探针。公共卫生相关性：我们的目标是确定能够调节SIAH-1蛋白酶体降解途径的化学物质。通过其作为泛素连接酶的活性，这种信号通路有助于调节各种细胞事件，包括细胞周期停滞，抑制肿瘤和凋亡。 SIAH-1途径的化学调节剂将提供强大的研究工具，用于研究与人类癌症发展，进展和对治疗耐药性有关的途径。