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

基本信息

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
项目类别：
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

项目摘要

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.

项目概要由蛋白精氨酸甲基转移酶 1 (PRMT1) 催化的蛋白精氨酸甲基化是目前作为抗癌治疗靶点以及肺、肾、神经退行性和心血管病理。迄今为止，大多数研究描述 PRMT1 介导的蛋白质甲基化的后果尚未考虑 PRMT1 活性改变的生化机制，这对于理解至关重要 PRMT1如何参与细胞稳态以及上述疾病的进展州，并制定治疗方案来控制 PRMT1 依赖性细胞事件。这该项目测试的两个假设是：1）寡聚状态和位点特异性的变化 PRMT1 蛋白的半胱氨酸氧化影响底物靶向和活性，2) PRMT1 中的半胱氨酸残基可用于创建/升级异构体特异性 PRMT1 抑制剂。在目标 1 中，将使用基于荧光的新型生物物理方法来表征完整细胞中 PRMT1 寡聚体形成的动态。战略性设计的变体以四聚体、二聚体或单体形式存在的 PRMT1 将用于识别结合和每种低聚物靶向的催化差异，有助于持续努力了解 PRMT1对其底物的分子识别规则。半胱氨酸磺酰化的影响 PRMT1 活性位点附近的残基也将被表征，从而允许开发 PRMT1 的稳定氧化和还原模拟物可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 提出，并有望应用于表征机制的长期目标调节体内 PRMT1 活性。在目标 2 中，使用靠近亲核半胱氨酸的策略将探索 PRMT1 的活性位点以实现腺苷衍生物的共价抑制。这该策略将是挽救缺乏异构体特异性的抑制剂的一种有价值的方法，但是否则可以很好地抑制 PRMT 活性。共价抑制剂代表了一种新颖的原理开发 PRMT1 药物和研究工具的途径。本科生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项目并接受实验设计和分析、笔记本保存、科学写作和演示。