Identification of Allsoteric Ligands for Hepatic Nuclear Factor 4-alpha

肝核因子 4-α 变构配体的鉴定

基本信息

批准号：
8421932
负责人：
FRAYDOON RASTINEJAD
金额：
$ 52.79万
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：
2012
资助国家：
美国
起止时间：
2012-12-05 至 2015-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8421932
关键词：
Affinity Architecture Binding Biochemical Biological Assay Blood Glucose Cataloging Catalogs Cells Complex Consensus DNA DNA Binding DNA Binding Domain Diabetes Mellitus Disease Dose Employee Strikes Family Fatty Acids Gene Targeting Genes Genetic Transcription Glucose Goals Hepatic Human Hypoglycemia Inherited Insulin Knowledge Length Libraries Ligand Binding Domain Ligands Link Liver Mutagenesis Mutate Mutation Neonatal Non-Insulin-Dependent Diabetes Mellitus Nuclear Nuclear Receptors Pancreas Pharmaceutical Chemistry Pharmaceutical Preparations Pharmacology Physiology Platelet Factor 4 Point Mutation Population Post-Translational Protein Processing Preclinical Drug Evaluation Property Proteins Resolution Response Elements Site Specificity Structure Testing Therapeutic Agents Time Work assay development base functional restoration glucose production high throughput screening human HNF4A protein member mutant patient population polypeptide receptor response scaffold screening small molecule transcription factor

项目摘要

DESCRIPTION (provided by applicant): The hepatocyte nuclear factor 4-alpha (HNF4) is a member of the nuclear receptor family of transcription factors. HNF4 controls genes in the liver and pancreas related to insulin and glucose production. Mutations in HNF4 cause Maturity Onset of Diabetes in Young 1 (MODY1), and hyperinsulinemic hypoglycemia (HH) in humans. Members of the nuclear receptor superfamily have a hydrophobic pocket inside their ligand binding domains (LBDs). Traditional drug screening strategies exploit these pockets, guided by known structures of isolated LBDs. The field has lacked high-resolution structures for full-length receptors, and therefore not considered the possibility of other druggable sites elsewhere in these proteins. We have obtained a complete crystal structure HNF4 homodimer, in a functionally revealing complex with its DNA response element. This structure unexpectedly reveals two sites at domain-domain junctions, where small molecules could bind. Our mutational studies, and previous knowledge of post-translational modifications, show these sites can allosterically regulate the DNA affinity of HNF4 . We propose a high-throughput screening campaign, together with confirmatory assays and mechanistic studies that exploit these new sites in HNF4 . Our goal is to identify potentiators of DNA binding that benefit certain MODY1 and HH disease populations where receptor-DNA binding is compromised by point mutations in HNF4 .

描述（由申请人提供）：肝细胞核因子4-α（HNF4）是转录因子的核受体家族的成员。 HNF4控制与胰岛素和葡萄糖产生有关的肝脏和胰腺中的基因。 HNF4中的突变会导致年轻1（Mody1）和人类高胰岛素低血糖（HH）的糖尿病的成熟发作。核受体超家族的成员在其配体结合结构域（LBD）内有一个疏水口袋。传统的药物筛查策略利用了这些口袋，以已知的LBD结构为指导。该领域缺乏全长受体的高分辨率结构，因此不认为这些蛋白质其他地方其他可吸毒位点的可能性。我们已经在功能透露的与其DNA响应元件的复合物中获得了完整的晶体结构HNF4同型二聚体。这种结构出乎意料地揭示了小分子可以结合的域域连接处的两个位点。我们的突变研究以及先前对翻译后修饰的知识表明，这些位点可以变构调节HNF4的DNA亲和力。我们提出了一项高通量筛查活动，以及确认性测定和机械研究，以利用HNF4中的这些新站点。我们的目标是鉴定有助于某些Mody1和HH疾病种群的DNA结合的增强剂，在HNF4中，受体DNA结合受到受体-DNA结合的损害。