Molecular mechanism of ABHD6 enzymatic activity in neurons

总结 脑中最丰富的内源性大麻素（eCB）2-花生四烯酸甘油（2-AG）可被两种 不同的酶：单酰基甘油脂肪酶（MAGL）和含有结构域的甘油三酸酯水解酶6（ABHD 6）。选择性 这些酶的抑制增加了不同神经元隔室、突触前和突触后2-AG的净水平。 和突触后。因此，这些酶的选择性抑制诱导不同的 2-AG信号传导的时空增强和独特的或协同的治疗益处。 我们最近收集的结果表明，ABHD 6水解额外的单酰基甘油底物比2- AG和ABHD 6抑制剂仅在高活性神经元中增加2-AG的净水平，这表明了一种新的 分子机制为了增加我们在结构和原子水平上对ABHD 6的机械理解， 我们开始了一项努力，现在已经成功地纯化了几种功能性ABHD 6蛋白构建体， 结构分析基于这一前提，我们提出了以下两个目标，这两个目标将决定： 目的1：ABHD 6酶活性的分子机制。 目的2：ABHD 6的结构与功能关系。 完成这项新的R21赠款提案中的工作大纲将全面了解 ABHD 6在神经系统疾病背景下控制脑中eCB信号传导中的作用。我们的长期 我们的目标是增加我们对ABHD 6在健康和患病大脑中所起作用的理解， 开发新的治疗方法，避免滥用和传统大麻素产生的副作用 激动剂

Summary The most abundant endocannabinoids (eCB) in brain, 2-arachidonoyl glycerol (2-AG), is inactivated by two distinct enzymes: monoacylglycerol lipase (MAGL) and hydrolase domain-contain 6 (ABHD6). The selective inhibition of these enzymes increases the net levels of 2-AG in different neuronal compartments, presynaptic and postsynaptic, respectively. Accordingly, selective inhibition of these enzymes induces different spatiotemporal enhancement of 2-AG signaling and distinct or synergistic therapeutic benefits. We recently gathered results showing that ABHD6 hydrolyzes additional monoacylglycerol substrates than 2- AG and that ABHD6 inhibitors increase the net levels of 2-AG only in highly activity neurons, suggesting a novel molecular mechanism. To increase our mechanistic understanding of ABHD6 at the structural and atomic levels, we initiated an effort and have now successfully purified several functional ABHD6 protein constructs ready for structural analysis. Based on this premise, we propose the following two aims that will determine the: Aim 1: Molecular mechanisms of ABHD6 enzymatic activity. Aim 2: Structure function relationship of ABHD6. Completion of the work outline in this new R21 grant proposal will provide a comprehensive understanding of the role of ABHD6 in controlling eCB signaling in brain within the context of neurological diseases. Our long-term goal is to increase our understanding of the role played by ABHD6 in healthy and diseased brain and help develop novel therapeutics that lack the potential for abuse and adverse effects produced by classic cannabinoid agonists.