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

• 批准号: 10033339
• 负责人: Edith C Glazer
• 金额: $ 34.18万
• 依托单位: UNIVERSITY OF KENTUCKY
• 依托单位国家: 美国
• 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/10033339
• 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; Estrogen receptor positive; 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

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.

摘要 CYP1B1在肝脏和健康组织中不存在或表达水平很低，而在 肿瘤，使其具有“万能肿瘤抗原”的称号。来自基础科学、临床和流行病学的证据 研究表明，CYP1B1参与了癌症的发生、发展和对多种药物的耐药性。 化疗药物。此外，在CYP1B1中发现了几个点突变，这些突变是 与其他疾病状态有关，特别是遗传性先天性青光眼和代谢紊乱。 然而，目前还没有已知的分子机制来解释突变、改变蛋白质水平、 而环境条件会影响这种酶的功能和功能障碍。我们假设细胞色素P1B1 亚细胞定位、寡聚、蛋白质：蛋白质组装、降解和小分子结合 由于特定的单核苷酸多态或环境特征而在癌细胞中发生改变。 CYP1B1从内质网到线粒体的运输，结合到多聚体状态，或产生 异常蛋白质：蛋白质组合可能导致蛋白质的破坏水平，从而导致新陈代谢 包括ROS和诱变剂在内的促进恶性进展的产品。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的活性和蛋白质的生命周期。单人 分子显微镜方法和荧光寿命显微镜成像将被用来获得高分辨率 关于活细胞中蛋白质组装和活性的信息。此外，这种酶的范式转换抑制剂 将通过两种不同的机制发挥作用：长时间滞留-时间协同抑制和 激活细胞色素P1B1的降解。CYP1B1蛋白水平和活性的降低应抑制其在 它可以促进癌症的进展，并有望促进各种化疗药物的疗效恢复。