Regulation of and Target Recognition by Protein Arginine Methyltransferase 1 (PRMT1)

蛋白质精氨酸甲基转移酶 1 (PRMT1) 的调节和目标识别

• 批准号: 10653465
• 负责人: JOAN M HEVEL
• 金额: $ 41.37万
• 依托单位: UTAH STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-05-01 至 2026-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10653465
• 关键词: Active Sites; Adenosine; Affect; Area; Arginine; Binding; Biochemical; Biochemistry; Biological; Biomedical Research; Cardiovascular Diseases; Cardiovascular Pathology; Catalysis; Cell Culture Techniques; Cell Cycle Regulation; Cell Extracts; Cell physiology; Cells; Chemistry; Chronic Disease; Communication; Cysteine; DNA Repair; Development; Disease; Disease Progression; Enzymes; Estrogen Receptors; Eukaryotic Cell; Event; Experimental Designs; Family; Fluorescence; Future; Goals; Grouping; Health; Histone H4; Histones; Homeostasis; Human; In Vitro; Inflammatory; Intervention; Kidney; Kidney Diseases; Kinetics; Lung; Lung diseases; Mammalian Cell; Mediating; Mediator; Medicine; Methylation; Modification; N,N-dimethylarginine; Nerve Degeneration; Neurodegenerative Disorders; Oxidative Stress; Pharmaceutical Preparations; Pharmacologic Substance; Play; Post-Translational Protein Processing; Progesterone Receptors; Protein Dynamics; Protein Inhibition; Protein Isoforms; Protein Methylation; Protein-Arginine N-Methyltransferase; Proteins; Regulation; Research; Research Personnel; Role; Signal Transduction; Site; Specificity; Spectrum Analysis; Stress; Testing; Training; Transcriptional Regulation; Translations; Treatment Protocols; United States National Institutes of Health; Universities; Utah; Variant; Viral Pathogenesis; Work; Writing; anti-cancer therapeutic; biophysical techniques; carcinogenesis; career; chemical reaction; design; dimer; dimethylarginine; drug development; epigenetic regulation; experimental analysis; graduate student; in vivo; inhibitor; interest; molecular recognition; monomer; non-histone protein; novel; oxidation; programs; protein function; small molecule; supportive environment; therapeutic target; tool; undergraduate research; undergraduate student

PROJECT SUMMARY Protein arginine methylation catalyzed by Protein arginine methyltransferase 1 (PRMT1) is of intense current interest as an anti-cancer therapeutic target, as well as a mediator of lung, kidney, neurodegenerative and cardiovascular pathologies. To date, the majority of studies characterizing the consequences of PRMT1-mediated protein methylation have not considered the biochemical mechanisms by which PRMT1 activity is altered, which is pivotal in understanding how PRMT1 participates in cellular homeostasis, the progression of the aforementioned disease states, and in developing treatment protocols to control PRMT1-dependent cellular events. The two hypotheses being tested in this project are: 1) changes in oligomeric state and site-specific cysteine oxidation of the PRMT1 protein affect substrate targeting and activity, and 2) that a cysteine residue in PRMT1 can be harnessed to create/upgrade isoform-specific PRMT1 inhibitors. In Aim 1, novel fluorescence-based biophysical methods will be used to characterize the dynamics of PRMT1 oligomer formation in intact cells. Strategically designed variants of PRMT1 which present as tetramers, dimers, or monomers will be used to identify binding and catalytic differences in the targeting by each oligomer, aiding in the ongoing effort to understand molecular recognition rules of PRMT1 for its substrates. The effect of sulfenylation at a cysteine residue near the active site of PRMT1 will also be characterized, allowing for the development of stable oxidized and reduced mimics of PRMT1 to be used as research tools in areas of human health affected by oxidative stress. The objectives in Aim 1 are built upon initial in vitro findings by the PI and are expected to apply to the long-term goal of characterizing mechanisms of regulating PRMT1 activity in vivo. In Aim 2, the strategy of using a nucleophilic cysteine near the active site of PRMT1 to enable covalent inhibition by adenosine derivatives will be explored. This strategy would be a valuable way to salvage inhibitors that show a lack of isoform-specificity, but otherwise perform well to inhibit PRMT activity. Covalent inhibitors represent a novel, rationale avenue for developing PRMT1 drugs and research tools. Undergraduate and graduate researchers with interests in future careers in biomedical research or medicine will be involved in the project and receive training in experimental design and analysis, notebook keeping, scientific writing and presentation.

项目摘要 蛋白质精氨酸甲基转移酶1（PRMT 1）催化的蛋白质精氨酸甲基化是一种新的生物学活性酶。 作为抗癌治疗靶点以及肺、肾 神经退行性疾病和心血管疾病。到目前为止，大多数研究 表征PRMT 1介导的蛋白质甲基化的后果尚未考虑 PRMT 1活性改变的生化机制，这是理解PRMT 1活性的关键。 PRMT 1如何参与细胞内稳态，上述疾病的进展 国家，并在开发治疗方案，以控制PRMT 1依赖性细胞事件。的 在这个项目中测试的两个假设是：1）寡聚状态和位点特异性的变化 PRMT 1蛋白的半胱氨酸氧化影响底物靶向和活性，以及2） PRMT 1中的半胱氨酸残基可用于产生/升级亚型特异性PRMT 1 抑制剂的在目标1中，将使用新的基于荧光的生物物理方法来表征 完整细胞中PRMT 1寡聚体形成的动力学。战略性设计的变体 作为四聚体、二聚体或单体存在的PRMT 1将用于鉴定结合和 每种寡聚体在靶向上的催化差异，有助于理解 PRMT 1对底物的分子识别规律。在半胱氨酸处的亚磺酰化的影响 PRMT 1活性位点附近的残基也将被表征，允许开发 PRMT 1的稳定氧化和还原模拟物用作人类领域的研究工具 健康受到氧化应激的影响。目标1中的目标建立在初步体外研究结果的基础上 由PI，并预计适用于表征机制的长期目标， 调节体内PRMT 1活性。在目标2中，使用亲核半胱氨酸接近目标2的策略是在目标2中使用亲核半胱氨酸。 将探索PRMT 1的活性位点以使腺苷衍生物能够共价抑制。这 策略将是挽救缺乏亚型特异性的抑制剂的有价值的方法，但 否则表现良好以抑制PRMT活性。共价抑制剂代表了一种新的， 开发PRMT 1药物和研究工具的途径本科和研究生 对生物医学研究或医学未来职业感兴趣的研究人员将参与 该项目并接受实验设计和分析，笔记本保存，科学 写作和演示。