A chemical biology approach to interrogate the zDHHC enzymes

药物研究的核心是发现和验证药物发现的新靶点。该项目以医疗保健技术为主题，与合成有机化学、计算化学和化学生物学的研究领域相结合，提供化学工具来询问zDHHC酶作为发现研究的目标。s -酰化是脂肪酸在半胱氨酸残基上的附着，调节多种蛋白质的活性，并由zDHHC家族酶催化。目前还没有化学工具可以用来理解这些酶的生物学特性。基于导师之间最近的合作，我们将使用化学生物学中的新兴技术为这类酶提供小分子探针。这项工作将分三个不同的阶段进行：第一阶段：zDHHC酶的制备、分离和纯化。这将在张伯伦实验室使用既定的技术进行。最初将制备三个不同的目标：hDHHC2催化结构域。hDHHC17催化域。hDHHC17锚蛋白重复结构域。第二阶段：与项目合作伙伴GSK合作。利用最近制备的光亲和片段文库，我们的酶将在这个550个成员文库中进行筛选。使用既定的筛选方案，将蛋白与单个光亲和片段孵育，然后在302 nm处照射。使用完整蛋白质谱分析将识别任何结合片段。后续分析将确认命中目标。阶段3：确定的片段命中将使用结构指导的迭代药物化学活动来改进活性。内部主屏幕将指导这项工作。这一阶段工作的目标是为每个检测目标提供亚微摩尔抑制剂。如果时间允许，后续的工作将寻找hDHHC2催化结构域和hDHHC17催化结构域之间的选择性。结果：预计这项工作将提供zDHHC酶家族的第一个抑制剂，这将使这些蛋白质的基本生物学研究成为可能。

At the core of pharmaceutical research is the discovery and validation of new targets for drug discovery. This project resides in the Healthcare Technologies theme, aligning with the research areas of synthetic organic chemistry, computational chemistry and the growth area of chemical biology to deliver chemical tools to interrogate zDHHC enzymes as targets for discovery research.S-Acylation, the attachment of fatty acids onto cysteine residues, regulates the activity of a wide range of proteins and is catalysed by the zDHHC family of enzymes. There are no chemical tools available to understand the biology of these enzymes. Based upon recent joint work between the supervisors we will use an emerging technology in chemical biology to deliver small molecule probes for this class of enzyme.The work will be performed in three distinct phases:Phase 1: The preparation, isolation and purification of zDHHC enzymes. This will be performed in the Chamberlain laboratory using established techniques. Initially three distinct targets will be prepared: hDHHC2 catalytic domain. hDHHC17 catalytic domain. hDHHC17 ankyrin repeat domain.Phase 2: This will be performed with project partners GSK. Using a recently prepared library of photoaffinity fragments our enzymes will be screened against this 550 member library. Using an established screening protocol protein will be incubated with individual photoaffinity fragments then irradiated at 302 nm. Analysis using intact protein mass spectrometry will identify any binding fragments. Follow up assays will confirm hit identification.Phase 3: Fragment hits identified will be elaborated using a structure guided iterative medicinal chemistry campaign to improve activity. In-house primary screens will guide this work. The goal of this phase of the work is to deliver a sub-micromolar inhibitor for each of the targets examined. Subsequent work if time permits will look for selectivity between the hDHHC2 catalytic domain and the hDHHC17 catalytic domain.Outcome: It is expected this work will deliver the first inhibitors of the zDHHC family of enzymes that will enable investigation of the fundamental biology of these proteins.