Development and Application of Synthetic Receptors for Methylated Lysine and Arginine for Methyltransferase Assays

用于甲基转移酶检测的甲基化赖氨酸和精氨酸合成受体的开发和应用

• 批准号: 1608333
• 负责人: Marcey Waters
• 金额: $ 47.51万
• 依托单位: University of North Carolina at Chapel Hill
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-09-01 至 2020-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1608333&HistoricalAwards=false
• 关键词: Development; Application; Synthetic; Receptors; Methylated

With this award, the Chemistry of Life Processes Program in the Chemistry Division is funding Dr. Marcey Waters from the University of North Carolina at Chapel Hill. While every cell in an individual person has the same genes, not all genes in a cell are expressed. Gene expression is the process by which genetic instructions are used to synthesize genetic products. Differences in gene expression lead to differentiation, such that a skin cell is different from a liver cell, for example. Research into understanding how genes are expressed is an important area because the factors that control gene expression can lead to many diseases. One factor that controls gene expression, but that is poorly understood, is a phenomenon called protein methylation, which involves the "tagging" of certain proteins with a molecule, here a methyl group. Studies indicate that incorrect methylation is a critical component in many diseases, particularly cancer, and that the processes that lead to methylation are potential medicinal targets. However, the ability to detect methylation is currently severely limited. Dr. Waters is developing new detection tools for rapidly sensing methylation, and determining when incorrect methylation occurs. This project is also supporting continued efforts to increase diversity in science fields and provide undergraduate and graduate student research opportunities for those from underrepresented groups.This research project is developing synthetic receptors as molecular sensors for methylated lysine and arginine, which are being used to characterize the enzymes that install and remove these post-translational modifications. Dysregulation of lysine and arginine methylation has been linked to a wide range of diseases, yet the tools available for sensing these modifications are limited and this has hindered advancements in the field. These approaches include the use of antibodies, which are expensive and have been shown to give false negatives; radioactive labeling, which is a low-throughput single-timepoint assay and requires specialized equipment and safety protocols; and mass spectrometry, which is time consuming and requires very sensitive, expensive equipment. The approach taken in this proposal is providing a rapid, real-time, high throughput, fluorogenic sensing method, which is enabling a significant advancement in the ability to characterize lysine and arginine methylation.

通过这一奖项，化学系的生命过程化学项目为北卡罗来纳大学教堂山分校的Marcey Waters博士提供了资金。虽然一个人体内的每个细胞都有相同的基因，但并不是一个细胞中的所有基因都得到了表达。基因表达是利用遗传指令合成基因产物的过程。例如，基因表达的差异会导致分化，比如皮肤细胞不同于肝细胞。了解基因是如何表达的研究是一个重要的领域，因为控制基因表达的因素可能导致许多疾病。控制基因表达的一个因素是一种被称为蛋白质甲基化的现象，它涉及到用一个分子(这里是甲基团)对某些蛋白质进行标记。研究表明，不正确的甲基化是许多疾病的关键组成部分，特别是癌症，而导致甲基化的过程是潜在的药物靶点。然而，目前检测甲基化的能力严重有限。沃特斯博士正在开发新的检测工具，用于快速检测甲基化，并确定何时发生错误的甲基化。该项目还支持继续努力增加科学领域的多样性，并为那些来自代表性不足群体的人提供本科生和研究生的研究机会。该研究项目正在开发合成受体，作为甲基化赖氨酸和精氨酸的分子传感器，这两种物质正被用来表征安装和移除这些翻译后修饰的酶。赖氨酸和精氨酸甲基化的失调与一系列疾病有关，但可用于检测这些修饰的工具有限，这阻碍了该领域的进展。这些方法包括使用昂贵的抗体，已被证明会产生假阴性；放射性标记，这是一种低通量的单一时间点分析，需要专门的设备和安全方案；以及质谱仪，这是耗时的，需要非常敏感、昂贵的设备。本提案中采用的方法提供了一种快速、实时、高通量的荧光传感方法，这使得表征赖氨酸和精氨酸甲基化的能力有了显著的进步。