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.

有了这个奖项，化学部门的化学过程计划是为北卡罗来纳大学教堂山分校的玛西·沃特斯博士提供资金。 虽然个体中的每个细胞都有相同的基因，但并非表达细胞中的所有基因。 基因表达是使用遗传指令合成遗传产品的过程。基因表达的差异导致分化，因此皮肤细胞与肝细胞不同。 研究理解基因如何表达是一个重要领域，因为控制基因表达的因素会导致许多疾病。 控制基因表达但知之甚少的一个因素是一种称为蛋白甲基化的现象，它涉及用分子的某些蛋白质的“标记”，这里是甲基。 研究表明，不正确的甲基化是许多疾病，尤其是癌症的关键成分，导致甲基化的过程是潜在的药用靶标。 但是，目前检测到甲基化的能力受到严重限制。 沃特斯博士正在开发新的检测工具，以快速感测甲基化，并确定何时发生不正确的甲基化。 该项目还支持持续努力，以增加科学领域的多样性，并为代表性不足的组的人提供本科和研究生的研究机会。本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，用于甲基化赖氨酸和精氨酸，这些传感器用于表征和删除这些变态后的酶。 赖氨酸和精氨酸甲基化的失调与多种疾病有关，但是可感知这些修饰的工具受到限制，这阻碍了该领域的进步。 这些方法包括使用抗体，这些抗体很昂贵，并且已被证明会产生错误的负面因素；放射性标签，这是一种低通量单个时间点测定，需要专门的设备和安全协议；和质谱，这很耗时，需要非常敏感，昂贵的设备。该提案中采用的方法提供了一种快速，实时，高通量的荧光感应方法，这使得能够表征赖氨酸和精氨酸甲基化的能力取得了显着进步。