Multidisciplinary approaches to characterise deacylating enzymes for therapeutic intervention

我们体内的细胞含有数千种不同的蛋白质，这些蛋白质对我们的组织和器官的正确结构和功能是必不可少的。蛋白质功能受到严格控制，以确保它们调控的独特和多样化的途径得到正确的协调，例如，通过添加某些化学基团-这一过程被称为翻译后修饰(PTM)。其中一种是“S酰化”，即一种被称为脂肪酸的化学基团可逆地连接到蛋白质上的特定位置。我们建议对S-酰化进行深入研究，因为这种形式的PTM与癌症和糖尿病等疾病以及精神分裂症、亨廷顿氏病和阿尔茨海默病等大脑疾病有关。一组从蛋白质中移除额外脂肪酸的蛋白质-称为“APT”酶-在S-酰化过程中至关重要，并支撑正常的细胞过程。细胞APT酶属于一个更大的酶家族，APT的总数比人们想象的要多样化得多，近年来发现了几种新的APT。在这个鲜为人知的研究领域，一个重要的突破是我最近发现了一种名为ABHD16A的蛋白质，它是一种全新的APT酶。ABHD16A与疼痛和炎症有关，我目前的一个研究项目是探索其作为治疗这些慢性疾病的药物靶点的潜力。此次奖学金旨在以我在APT发现科学和更广泛的S-酰化领域的成熟专业知识为基础，揭示调节动态S-酰化途径的细胞APT，并确定抑制其活性的分子。这些发现对于加快我们对S酰化在与人类疾病相关的细胞通路中的作用的理解是至关重要的，并使我们能够探索APT作为新的治疗干预措施的靶向的潜力。为了实现这些目标，我已经建立了一个合作者网络，他们是化学生物学、化学蛋白质组学和分子建模方面的国际专家，我将使用一套我经验丰富的最新开发的尖端技术。与尼克·汤金森教授领导的斯特拉斯克莱德大学的有机化学家合作，我将利用我最近开发的化学生物学工具，准确测量细胞中APT表达水平变化后蛋白质S酰化水平的变化。操纵APT酶表达的影响将在华威大学的蛋白质组学研究技术平台上与化学蛋白质组学专家Andrew Bottrill博士和同事一起，使用现有最先进和最灵敏的质谱学方法进行定量测量。由克里斯·雷诺兹教授领导的考文垂大学计算化学专家将利用生物信息学、比较模型和分子动力学模拟来生成高质量的结构，以了解APT酶的功能并开发调节APT酶活性的化合物。最终，这项研究计划将阐明APT酶如何协调基本的S-酰化过程，查明在特定人类疾病中存在缺陷的机制，并为发现治疗一系列威胁全球人类健康的疾病的新药确定新的治疗靶点。

The cells in our body contain thousands of different proteins that are essential for the correct structure and functionof our tissues and organs. Protein function is tightly controlled to ensure that the unique and diverse pathwaysthat they regulate are correctly coordinated, for example, through the addition of certainchemical groups - a process called Post-Translational Modification (PTM). One such PTM is 'S-acylation' in which chemical groups called fatty-acids are reversibly attached to specific sites on proteins. We propose an in-depth study of S-acylation as this form of PTM is linked to diseases such as cancer and diabetes, and brain disorders such as schizophrenia, Huntington's disease and Alzheimer's disease.A group of proteins that remove added fatty acids from proteins - called "APT" enzymes - are critical to this S-acylationprocess and underpin normal cellular processes. Cellular APT enzymes belong to a much larger enzyme familyand the total number of APTs is more diverse than previously thought, with several novel APTs having been identified inrecent years. One such important breakthrough in this poorly-understood area of research was my recent discovery of aprotein called 'ABHD16A' as an entirely new APT enzyme. ABHD16A is associated with pain and inflammation and one ofmy current research projects is exploring its potential as a drug target to treat these chronic diseases.This fellowship aims to build on my proven expertise in APT discovery science and in the wider S-acylation field to revealcellular APTs that regulate dynamic S-acylation pathways and identify molecules that inhibit their activity. Thesediscoveries are essential to expedite our understanding of the role of S-acylation in cellular pathways linked to humandiseases and to enable us to explore the potential of APTs to be targeted in new therapeutic interventions.To address these aims, I have put in place the support of a network of collaborators who are international experts inchemical biology, chemical proteomics and molecular modelling and I will use a comprehensive suite of recently-developedcutting-edge techniques that I am experienced in. In collaboration with organic chemists at the University of Strathclyde ledby Prof Nick Tomkinson, I will utilise chemical-biology tools that I have recently developed to accurately measure changesin the level of protein S-acylation following alterations in the level of APT expression in cells. The effects of manipulatingthe expression of APT enzymes will be measured quantitatively at the Proteomics Research Technology Platform, University of Warwick, with Dr Andrew Bottrill andcolleagues who are experts in chemical proteomics, using the most advanced and sensitive mass spectrometry processesavailable. Experts in computational chemistry at Coventry University, led by Prof Chris Reynolds, will use bioinformatics, comparative modelling and molecular dynamic simulations to generate high quality structures for the purpose of understanding function and for the development of compounds that modulate the activity of APT enzymes.Ultimately, this research programme will elucidate how APT enzymes coordinate essential S-acylation processes, pinpointing the mechanisms that are faulty in specific human diseases and identifying new therapeutic targets for thediscovery of novel drugs to treat a range of diseases that threaten global human health.