Interrogating Protein Methyltransferases with Integrated Approaches

项目摘要/摘要：蛋白质甲基化是一种表观遗传事件，由>60 人蛋白质甲基转移酶（PMT）。因为许多PMT相关的甲基化事件是 在指定的细胞环境下，我们对传统方法的理解是不可见的， PMT的表观遗传作用非常有限。光电倍增管的选择性扰动是至关重要的， 阐明PMT的疾病特异性作用并制定新的治疗策略。背景下 这些技术和知识差距，在过去的五年里，我们的实验室专注于 开发新的技术并实施它们来准确地注释非组蛋白 甲基化事件和下游分子机制的PMT。同时，我们的实验室 也投入了大量的精力来研究PMT涉及的原子机制 催化和利用它们合理地开发PMT抑制剂通过不同的相互作用模式。 在这里，我们计划扩展这些发现。我们设想表征非组蛋白甲基化事件 与转录激活相关的综合方法，包括化学标记， Criminal/Cas9编辑。我们将通过SAM竞争性或共价结合的方式开发高质量的PMT抑制剂， 模式，特别是考虑到PMT的动态构象景观。的 完成这一建议将明确揭示多种PMT的分子机制， 相关的细胞环境，并提供高质量的PMT抑制剂的扰动。附加 我们的功能注释和通道标识策略的影响源于它们的一般 适用于其他甲基转移酶。

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.