Synthetic probes of histidine phosphorylation: new reagents for systems biology and proteomics

组氨酸磷酸化合成探针：系统生物学和蛋白质组学新试剂

• 批准号: EP/I013083/1
• 负责人: Michael Webb
• 金额: $ 12.86万
• 依托单位: University of Leeds
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2011
• 资助国家: 英国
• 起止时间: 2011 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=EP%2FI013083%2F1
• 关键词: Synthetic; probes; histidine; phosphorylation; new

DNA sequencing has revealed that the human genome comprises only 30000 distinct proteins. That such a relatively small number of proteins can combine to form a human being is due to the vast number of ways in which they can be modified to control their function and behaviour. The dominant method by which this occurs is protein phosphorylation, a vast body of research in the last fifty years (now comprising several thousand publications/year) has demonstrated how modification of the amino acids serine, threonine and tyrosine control the physiology of the cell and whole organisms. Errors in these control and signalling pathways are implicated in many human diseases including cancer.There are twenty natural amino acids in total and many others can also be phosphorylated. We are interested in exploring the biochemistry of phosphorylated histidine which has been little studied to date. N-linked Phosphohistidine is less stable than its O-linked counterparts and it has not therefore been possible to acquire antibodies capable of identifying this modification. This, in turn, means that it has been impossible to assess how prevalent the modification is in cells and how it is involved in regulating complex biochemical processes. In this project we will generate the first class of such reagents thereby enabling a step change in research in this area.We will synthesize a range of stable analogues of phosphohistidine and incorporate these into short peptides. We will then evaluate the analogues by measuring how well they bind to a protein known to recognise and modify phosphohistidine-containing proteins. Following this we will screen libraries of peptide aptamers for aptamers which specifically recognise the analogue and thereby phosphohistidine. Such a tool will then be applied to the assay of other biological systems.

脱氧核糖核酸测序显示，人类基因组只由30000种不同的蛋白质组成。如此少量的蛋白质能够结合在一起形成人类，是因为它们可以通过多种方式进行修饰，以控制它们的功能和行为。发生这种情况的主要方法是蛋白质磷酸化，在过去的50年里，大量的研究(现在包括每年数千篇论文)已经证明了氨基酸丝氨酸、苏氨酸和酪氨酸的修饰如何控制细胞和整个生物体的生理。这些控制和信号通路的错误与包括癌症在内的许多人类疾病有关。总共有20种天然氨基酸，还有许多其他氨基酸也可以被磷酸化。我们对探索磷酸化组氨酸的生物化学很感兴趣，到目前为止，对它的研究还很少。N-连接的磷酸组氨酸比其O-连接的对应物更不稳定，因此不可能获得能够识别这种修饰的抗体。这反过来意味着无法评估这种修饰在细胞中有多普遍，以及它是如何参与调控复杂的生化过程的。在这个项目中，我们将生产第一类此类试剂，从而使这一领域的研究发生阶段性变化。我们将合成一系列稳定的磷酸组氨酸类似物，并将它们合并到短肽中。然后，我们将通过测量它们与一种已知识别和修饰含磷酸组氨酸的蛋白质的结合情况来评估这些类似物。在此之后，我们将筛选肽适配子文库，寻找能够识别类似物从而识别磷酸组氨酸的适配子。这样的工具将被应用于其他生物系统的分析。

项目成果

Strain-promoted reaction of 1,2,4-triazines with bicyclononynes.

• DOI: 10.1002/chem.201502397
• 发表时间: 2015-10-05
• 期刊: Chemistry (Weinheim an der Bergstrasse, Germany)
• 作者: Horner KA;Valette NM;Webb ME
• 通讯作者: Webb ME

Evaluation of the interaction between phosphohistidine analogues and phosphotyrosine binding domains.

• DOI: 10.1002/cbic.201402090
• 发表时间: 2014-05-26
• 期刊: CHEMBIOCHEM
• 影响因子: 3.2
• 作者: McAllister, Tom E.;Horner, Katherine A.;Webb, Michael E.
• 通讯作者: Webb, Michael E.

Efficient N-terminal labeling of proteins by use of sortase.

• DOI: 10.1002/anie.201204538
• 发表时间: 2012-09-10
• 期刊: Angewandte Chemie (International ed. in English)
• 作者: Williamson, Daniel J;Fascione, Martin A;Turnbull, W Bruce
• 通讯作者: Turnbull, W Bruce

Prospects for stable analogues of phosphohistidine.

• DOI: 10.1042/bst20130071
• 发表时间: 2013-08
• 期刊: Biochemical Society transactions
• 影响因子: 3.9
• 作者: Tom E. McAllister;J. Hollins;M. Webb