Role of small molecule interactions and multiprotein assemblies in CYP1B1 disease-associated function and dysfunction

小分子相互作用和多蛋白组装在 CYP1B1 疾病相关功能和功能障碍中的作用

基本信息

批准号：
10242829
负责人：
Edith C Glazer
金额：
$ 36.62万
依托单位：
UNIVERSITY OF KENTUCKY
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-09-01 至 2024-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10242829
关键词：
3-Dimensional Affect Aromatase Aromatase Inhibitors Basic Science Binding Biology CYP17A1 gene CYP1B1 gene Cell Line Cell physiology Cells Chemotherapy-Oncologic Procedure Chimeric Proteins Cisplatin Clinical Research Conflict (Psychology)Cytochrome P450 Cytochrome a Data Disease Dose Drug resistance Environment Environmental Risk Factor Enzyme Inhibitor Drugs Enzymes Functional disorder Generations Genetic Genetic Polymorphism Goals Half-Life Heme Hemeproteins Homo Hormones Hydrophthalmos Hypoxia Immune response In Vitro Inherited Iron Liver Malignant - descriptor Malignant Neoplasms Malignant neoplasm of prostate Measurement Medical Membrane Metabolic Metabolic Diseases Methods Microscopy Mitochondria Modeling Molecular Molecular Conformation Mutagens Mutation Mycoses Nucleotides Paclitaxel Patients Pharmacology Phase I Clinical Trials Phenotype Photobleaching Physiologic pulse Play Point Mutation Population Population Distributions Proteins Proteolysis Resistance Resolution Role Scanning Single Nucleotide Polymorphism Testing Thermodynamics Time Tissues Tumor Antigens Vesicle Work cancer cell cancer initiation chemotherapy enzyme activity epidemiology study experimental study fluorescence lifetime imaging improved in vivo inhibitor/antagonist knock-down malignant breast neoplasm mitochondrial membrane nanoscale novel overexpression protein degradation protein protein interaction residence response restoration single molecule small molecule small molecule inhibitor trafficking tumor tumor progression

项目摘要

Abstract CYP1B1 is absent or expressed at very low levels in the liver and healthy tissues while being overexpressed in tumors, giving it the title of “universal tumor antigen”. Evidence from basic science, clinical, and epidemiological studies demonstrate that CYP1B1 is involved in cancer initiation, progression, and resistance to a wide range of chemotherapeutics. In addition, several point mutations have been discovered within CYP1B1 that are associated with other disease states, particularly hereditary congenital glaucoma and metabolic disorders. However, there is currently no known molecular mechanism that explains how mutations, altered protein levels, and environmental conditions impact the function and dysfunction of this enzyme. We hypothesize that CYP1B1 subcellular localization, oligomerization, protein:protein assembly, degradation, and small molecule binding are altered in cancer cells as a result of specific single nucleotide polymorphisms or environmental features. Trafficking of CYP1B1 from the ER to the mitochondria, association into multimeric states, or generation of abnormal protein:protein assemblies could result in damaging levels of the protein and, consequently, metabolic products including ROS and mutagens that facilitate malignant progression. This work will demonstrate how the protein:protein interactions of a cytochrome P450 enzyme impacts enzyme activity and protein lifecycle. Single molecule microscopy methods and fluorescence lifetime microscopy imaging will be used to gain high resolution information on protein assembly and activity in live cells. Additionally, paradigm-shifting inhibitors of this enzyme will be developed that act though two distinct mechanisms: long-residence-time coordinative inhibition, and activation of CYP1B1 degradation. Reduction of CYP1B1 protein levels and activity should suppress its role in cancer progression, and is anticipated to facilitate restoration of efficacy for a variety of chemotherapeutics.

抽象的 CYP1B1在肝脏和健康组织中不存在或表达非常低的水平肿瘤，赋予其标题为“通用肿瘤抗原”。来自基础科学，临床和流行病学的证据研究表明，CYP1B1参与了癌症的开始，进展和对广泛的抗药性化学治疗学。此外，在CYP1B1中发现了几个点突变与其他疾病状态，特别是遗传性先天性青光眼和代谢疾病有关。但是，目前尚无已知的分子机制来解释突变，改变蛋白质水平，如何改变，环境条件会影响该酶的功能和功能障碍。我们假设CYP1B1 亚细胞定位，低聚，蛋白质：蛋白质组装，降解和小分子结合是由于特定的单核苷酸多态性或环境特征而改变了癌细胞的改变。将CYP1B1从ER到线粒体的贩运，与多媒体状态相关或产生异常蛋白质：蛋白质组件可能导致蛋白质的破坏水平，因此代谢包括促进恶性进展的ROS和诱变者在内的产品。这项工作将展示如何蛋白质：细胞色素P450酶的蛋白质相互作用会影响酶活性和蛋白质生命周期。单身的分子显微镜方法和荧光寿命显微镜成像将用于获得高分辨率有关活细胞中蛋白质组装和活性的信息。另外，该酶的范式转移抑制剂将开发出两种不同机制的作用：长期居住时间的坐标抑制和 CYP1B1降解的激活。 CYP1B1蛋白水平和活性的降低应抑制其在癌症的进展，预计有助于恢复各种化学治疗剂的效率。