A chemical biology approach to interrogate the zDHHC enzymes

询问 zDHHC 酶的化学生物学方法

• 批准号: 2267309
• 金额: --
• 依托单位: University of Strathclyde
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2019
• 资助国家: 英国
• 起止时间: 2019 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2267309
• 关键词: chemical; biology; approach; interrogate; zDHHC

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.

药物研究的核心是发现和验证药物发现的新靶点。该项目属于医疗保健技术主题，与合成有机化学、计算化学和化学生物学的增长领域相结合，提供化学工具来询问zDHHC酶作为发现研究的目标。S-酰化，脂肪酸连接到半胱氨酸残基上，调节各种蛋白质的活性，并由zDHHC酶家族催化。目前还没有化学工具来了解这些酶的生物学特性。基于近期双方主管的共同工作，我们将利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Chamberlain实验室使用既定技术进行。最初将准备三个不同的靶点：hDHHC 2催化结构域。hDHHC 17催化结构域。第2阶段：这将与项目合作伙伴GSK一起进行。使用最近制备的光亲和片段库，我们的酶将针对这550个成员库进行筛选。使用已建立的筛选方案，将蛋白质与单个光亲和片段一起孵育，然后在302 nm下照射。使用完整蛋白质质谱法的分析将鉴定任何结合片段。后续检测将确认命中鉴定。第3阶段：将使用结构指导的迭代药物化学活动来详细说明鉴定的片段命中，以提高活性。内部主要屏幕将指导这项工作。这一阶段工作的目标是为每个检查的目标提供亚微摩尔抑制剂。如果时间允许，后续工作将寻找hDHHC 2催化结构域和hDHHC 17催化结构域之间的选择性。结果：预计这项工作将提供zDHHC家族酶的第一个抑制剂，这将使这些蛋白质的基础生物学研究成为可能。