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.

DNA测序表明，人类基因组仅包含30000种不同的蛋白质。如此少量的蛋白质可以结合起来形成人类，这是由于可以修改它们以控制其功能和行为的大量方式。发生这种情况的主要方法是蛋白质磷酸化，这是过去五十年来的大量研究（现在包括数千份出版物/年）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学。这些对照和信号通路中的错误与包括癌症在内的许多人类疾病有关。总共有二十种天然氨基酸，许多其他氨基酸也可以磷酸化。我们有兴趣探索迄今为止很少研究的磷酸化组氨酸的生物化学。 N-连接的磷酸化酶氨酸的稳定性不如其O-Link的对应物，因此不可能获得能够识别这种修饰的抗体。反过来，这意味着无法评估细胞中的修饰的普遍性以及如何参与调节复杂的生化过程。在这个项目中，我们将生成此类试剂的第一类，从而实现该领域的研究的步骤变化。我们将合成一系列磷酸磷酸组织的稳定类似物，并将其掺入短肽中。然后，我们将通过测量它们与已知识别和修饰含磷酸组织的蛋白质的蛋白质结合的结合来评估类似物。此后，我们将筛选适体的肽适体的文库，这些肽适体特有特殊识别模拟并因此磷酸化组氨酸。然后将这种工具应用于其他生物系统的测定法。

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.

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

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