Molecular imaging approaches to interrogate mammalian signaling by lysine acylation

项目摘要 人类蛋白质组中的数千种蛋白质都受到可逆赖氨酸的修饰 乙酰化，现在被认为是多种过程的关键调节剂， 表观遗传学和新陈代谢。赖氨酸的表达和活性之间的谨慎平衡 乙酰转移酶（KAT）和赖氨酸脱乙酰酶（KDAC）维持细胞的乙酰组。 我们的长期目标是对赖氨酸乙酰化是如何 控制以及标记如何影响细胞状态。我们目前的重点是开发一个新的类 小分子荧光化学工具，其报告活体中赖氨酸脱乙酰化活性 具有空间分辨率的细胞。我们将部署这一新的化学工具系列， 假设KDAC信号传导部分是由亚细胞分布介导的， 获得底物和局部辅因子。这一假设可以帮助解释一些 与赖氨酸乙酰化相关的模棱两可的结果已经观察到，作为生物 特定KDAC同种型的调节的结果可以被细胞中的其他同种型所掩盖。 细胞类型或疾病特异性方式。我们假设了解赖氨酸 调节乙酰化的目的是监测KAT和KDAC活性的总量， 时空分辨率我们现在主要的生物学兴趣是KDAC的作用 细胞核外，特别是线粒体和细胞质中， 代谢和乳腺癌背景下的机制研究。

Project Summary Thousands of proteins in the human proteome are subjected to modification by reversible lysine acetylation, which is now recognized as a key regulator of diverse processes such as epigenetics and metabolism. The careful balance between the expression and activity of lysine acetyl transferases (KATs) and lysine deacetylases (KDACs) maintains the acetylome of a cell. Our long-term goal is to develop a mechanistic understanding of how lysine acetylation is controlled and how the mark effects cell state. Our current focus is to develop a new class of small molecule fluorescent chemical tools that report on lysine deacetylation activities in living cells with spatial resolution. We will deploy this new family of chemical tools to test the hypothesis that KDAC signaling is in part mediated by subcellular distribution, which controls access to substrates and local cofactors. This hypothesis could help explain some of the ambiguous results associated with lysine acetylation that have been observed, as the biological consequence of modulation of a specific KDAC isoform may be masked by other isoforms in a cell-type or disease specific manner. We postulate that a key to understanding how lysine acetylation is regulated is to monitor the overall amounts of KAT and KDAC activities with spatial-temporal resolution. Our primary biological interests right now deal with roles of KDACs outside of the nucleus, in particular in the mitochondria and the cytoplasm, while pursing mechanistic studies in the context of metabolism and breast cancer.