Interrogating Protein Methyltransferases with Integrated Approaches

用综合方法研究蛋白质甲基转移酶

• 批准号: 10436329
• 负责人: Minkui Luo
• 金额: $ 79.02万
• 依托单位: SLOAN-KETTERING INST CAN RESEARCH
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-06-01 至 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10436329
• 关键词: Biochemical; Biological Process; Biology; Catalysis; Chemicals; Disease; Epigenetic Process; Event; Human; Knowledge; Label; Laboratories; Lead; Methods; Methylation; Methyltransferase; Molecular; Molecular Conformation; Protein Methylation; Protein Methyltransferases; Role; Technology; Transcriptional Activation; inhibitor; new technology; novel therapeutic intervention; novel therapeutics; stem; tool

Project Summary/Abstract: Protein methylation is an epigenetic event that is regulated by >60 human protein methyltransferases (PMTs). Because many PMT-associated methylation events are invisible for conventional methods under designated cellular settings, our understanding of epigenetic roles of PMTs is very limited. Selective perturbation of PMTs is of critical need towards elucidating disease-specific roles of PMTs and formulating new therapy strategies. In the context of these technology and knowledge gaps, in the past five years, our laboratory has focused on developing novel technologies and implementing them to accurately annotate nonhistone methylation events and downstream molecular mechanisms of PMTs. Meanwhile, our laboratory has also devoted significant efforts to examine atomistic mechanisms underlying PMT-involved catalysis and leverage them to rationally develop PMT inhibitors via distinct modes of interaction. Here we plan to expand these findings. We envision characterizing nonhistone methylation events associated with transcription activation with integrated approaches including chemical labeling and Crisper/Cas9 editing. We will develop high-quality PMT inhibitors via SAM-competitive or covalent modes, in particular with consideration of dynamic conformational landscapes of PMTs. The completion of this proposal will unambiguously reveal molecular mechanisms of multiple PMTs in relevant cellular contexts and deliver high-quality PMT inhibitors for perturbation. The additional impact of our function-annotating and inhibitor-identifying strategies stems from their general applicability to other methyltransferases.

项目摘要/摘要：蛋白质甲基化是受&gt；60调控的表观遗传事件。 人蛋白甲基转移酶(PMTs)。因为许多与PMT相关的甲基化事件 在指定的蜂窝环境下，常规方法是不可见的，我们对 PMTs的表观遗传学作用非常有限。对PMTS的选择性扰动是迫切需要的 阐明PMTs的疾病特异性作用并制定新的治疗策略。在…的背景下 这些技术和知识的差距，在过去的五年里，我们的实验室一直专注于 开发新技术并实施以准确注释非组蛋白 PMTs的甲基化事件和下游分子机制。与此同时，我们的实验室 还投入了大量的精力来检查PMT参与的原子论机制 催化并利用它们通过不同的相互作用模式合理地开发PMT抑制剂。 在这里，我们计划扩大这些发现。我们设想了非组蛋白甲基化事件的特征 与转录激活相关的集成方法，包括化学标记和 CRISPER/Cas9编辑。我们将通过SAM开发高质量的PMT抑制剂-竞争或共价 模式，特别是考虑到PMT的动态构象景观。这个 这一建议的完成将明确揭示多种PMTs的分子机制。 相关的细胞环境，并提供高质量的PMT抑制剂用于干扰。附加的 我们的功能注释和抑制物识别策略的影响来自于它们的一般性 对其他甲基转移酶的适用性。