Mapping the acylated proteome: a chemical genetic approach

绘制酰化蛋白质组图谱：化学遗传学方法

• 批准号: BB/D02014X/1
• 负责人: Edward Tate
• 金额: $ 92.3万
• 依托单位: Imperial College London
• 依托单位国家: 英国
• 项目类别: Fellowship
• 财政年份: 2006
• 资助国家: 英国
• 起止时间: 2006 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FD02014X%2F1
• 关键词: Mapping; acylated; proteome; chemical; genetic

With the completion of several important genome projects, such as the Human Genome Project, we now have the basic code describing the proteins made by cells which carry out the essential processes of life. However, this is only the beginning of our journey towards fully understanding how cells function: the focus of research has now moved to unravelling the role of each protein in a cell, and when and where it is carried out. To further complicate the story, the structure of a protein is frequently modified after its initial creation (its 'translation' from the genetic code) by other proteins called enzymes, a process termed 'post-translational modification'. These modifications can have critically important functions in the cell; for example, they are involved in our immune response to disease and the mechanism of infection by viruses, and defective modification is implicated in diseases such as cancer. Drugs which target the post-translational enzymes in pathogenic organisms such as fungi and parasites (for example, Plasmodium falciparum, the single-celled parasite which causes malaria) are under development, and could make a major impact on our ability to control these diseases. However, due to the complexity of the total set of thousands of different proteins in a cell (dubbed the 'proteome'), it has proven difficult to identify which proteins are modified, and what effect the modification has on a protein's function. Identifying the modified proteome is a key step towards understanding the role of post-translational modification, and to discover what effects (and side-effects) the drugs mentioned above may have in the cell. The proposed research aims to address this challenge by integrating recent discoveries in organic chemistry with cutting-edge whole-proteome analysis. We aim to exploit the cell's own enzymes to introduce a chemical 'tag' into proteins which are modified by a process termed 'acylation'. The tags are designed so as not to disrupt the cell's normal function, and we can very selectively 'capture' and purify the tagged acyl proteins with specially designed reagents. This will allow us to hugely increase the sensitivity of whole-proteome analysis towards acyl modifications, revealing hitherto unidentified acyl proteins and opening up new lines of investigation into the role of acylation in cells. Later, we intend to exploit this tag and capture methodology to analyse the effect of drugs on the modified proteome in single-celled parasites, and even to enable us to see modified proteins under a microscope as they move around the cell. This work will bring together chemists at Imperial College London and biologists at the University of York in an 'interdisciplinary' collaboration / that is, at the interface between the traditional disciplines of Chemistry and Biology.

随着几个重要的基因组计划的完成，如人类基因组计划，我们现在有了描述细胞制造的蛋白质的基本代码，这些蛋白质执行生命的基本过程。然而，这只是我们全面了解细胞功能的开始：研究的重点现在已经转移到解开细胞中每种蛋白质的作用，以及何时何地进行。为了进一步复杂化的故事，蛋白质的结构经常被修改后，其最初的创造（其'翻译'从遗传密码）由其他蛋白质称为酶，一个过程称为'翻译后修饰'。这些修饰在细胞中具有至关重要的功能;例如，它们参与了我们对疾病的免疫反应和病毒感染的机制，而缺陷修饰与癌症等疾病有关。针对真菌和寄生虫（例如，恶性疟原虫，导致疟疾的单细胞寄生虫）等病原生物中的翻译后酶的药物正在开发中，可能会对我们控制这些疾病的能力产生重大影响。然而，由于细胞中数千种不同蛋白质（称为“蛋白质组”）的复杂性，已经证明很难识别哪些蛋白质被修饰，以及修饰对蛋白质功能的影响。鉴定修饰的蛋白质组是了解翻译后修饰的作用以及发现上述药物可能对细胞产生哪些影响（和副作用）的关键一步。拟议的研究旨在通过将有机化学的最新发现与尖端的全蛋白质组分析相结合来应对这一挑战。我们的目标是利用细胞自身的酶将化学“标签”引入蛋白质中，这些蛋白质通过称为“酰化”的过程进行修饰。标签的设计是为了不破坏细胞的正常功能，我们可以非常有选择性地“捕获”和纯化标记的酰基蛋白质与特殊设计的试剂。这将使我们能够极大地提高全蛋白质组分析对酰基修饰的灵敏度，揭示迄今为止尚未识别的酰基蛋白，并开辟新的研究路线，研究酰基化在细胞中的作用。后来，我们打算利用这种标签和捕获方法来分析药物对单细胞寄生虫中修饰蛋白质组的影响，甚至使我们能够在显微镜下看到修饰蛋白质在细胞中移动。这项工作将汇集化学家在帝国理工学院伦敦和生物学家在约克大学在一个“跨学科”的合作/也就是说，在化学和生物学的传统学科之间的接口。

项目成果

Mutational Locally Enhanced Sampling (MULES) for quantitative prediction of the effects of mutations at protein-protein interfaces

• DOI: 10.1039/c2sc00895e
• 发表时间: 2012-01-01
• 期刊: CHEMICAL SCIENCE
• 影响因子: 8.4
• 作者: Bradshaw, Richard T.;Aronica, Pietro G. A.;Gould, Ian R.
• 通讯作者: Gould, Ian R.

Diverse modes of binding in structures of Leishmania major N-myristoyltransferase with selective inhibitors.

• DOI: 10.1107/s2052252514013001
• 发表时间: 2014-07-01
• 期刊: IUCrJ
• 影响因子: 3.9
• 作者: Brannigan JA;Roberts SM;Bell AS;Hutton JA;Hodgkinson MR;Tate EW;Leatherbarrow RJ;Smith DF;Wilkinson AJ
• 通讯作者: Wilkinson AJ

Selective inhibitors of protozoan protein N-myristoyltransferases as starting points for tropical disease medicinal chemistry programs.

• DOI: 10.1371/journal.pntd.0001625
• 发表时间: 2012
• 期刊: PLoS neglected tropical diseases
• 影响因子: 3.8
• 作者: Bell AS;Mills JE;Williams GP;Brannigan JA;Wilkinson AJ;Parkinson T;Leatherbarrow RJ;Tate EW;Holder AA;Smith DF
• 通讯作者: Smith DF