Illuminating serine hydrolase activity

 描述（由申请人提供）：许多脂质及其衍生物是细胞受体的有效配体。脂质第二信使的作用受限制这些生物活性分子的寿命和作用范围的脂肪酶和相关酶的调节。这些水解反应的产物本身可以是第二信使和其他生物活性分子的前体。例如，丝氨酸水解酶单酰基甘油脂肪酶（MAGL）负责限制内源性大麻素2-花生四烯酸酰甘油（2-AG）的寿命和作用范围，并负责产生脂肪酸产物花生四烯酸。这些酶的抑制剂是用于治疗疼痛、焦虑、成瘾、炎症和其中炎症或脂质调节起中心作用的无数疾病的潜在治疗剂。然而，目前可用于评估丝氨酸水解酶活性和抑制的工具主要限于体外使用。在这项资助中，我们将开发丝氨酸水解酶的发光传感器（目的1），并将其应用于活细胞中丝氨酸水解酶活性的成像和这类酶的新型抑制剂的鉴定（im 2）。最后，我们将在体内对丝氨酸水解酶活性进行实时成像，以确定这些酶在何处以及何时活跃，以及药物泵的作用如何影响其功能的小分子抑制剂的组织分布（目的3）。

 DESCRIPTION (provided by applicant): Many lipids and their derivatives are potent ligands for cellular receptors. The action of lipid second messengers is regulated by lipases and related enzymes that limit the lifetime and sphere of action of these bioactive molecules. The products of these hydrolytic reactions can themselves be second messengers and precursors to other bioactive molecules. For example, the serine hydrolase monoacylglycerol lipase (MAGL) is responsible for limiting the lifetime and sphere of action of the endocannabinoid 2-arachidonoylglycerol (2-AG) and for generating the fatty acid product, arachidonic acid. Inhibitors of these enzymes are potential therapeutics for the treatment of pain, anxiety, addiction, inflammation, and a myriad of diseases where inflammation or lipid regulation play central roles. However, the tools currently available for the evaluation of serine hydrolase activity and inhibition are primarily limited to use in vitro. In this grant, we will develop luminogenic sensors for serine hydrolases (Aim 1), and apply them to the imaging of serine hydrolase activity in live cells and identification of novel inhibitors for this class of enzymes (im 2). Finally, we will perform real-time imaging of serine hydrolase activity in vivo, to determine where and when these enzymes are active, and how the action of drug pumps affect the tissue distribution of small molecule inhibitors of their function (Aim 3).