Tag & Charge A new approach to simultaneously enrich and enhance phosphoproteome analysis

标签

• 批准号: BB/I012354/1
• 负责人: William Griffiths
• 金额: $ 36.37万
• 依托单位: Swansea University
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2012
• 资助国家: 英国
• 起止时间: 2012 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FI012354%2F1
• 关键词: Tag; Charge; new; approach; simultaneously

Reversible protein phosphorylation has turned out to be a general control mechanism involved in almost all aspects of cellular process. The significance and complexity of reversible protein phosphorylation can be assessed from the fact that there are at least 518 protein kinases and more than 100 protein phosphatases present in the human genome, and it is estimated that one third of proteins are phosphorylated in a given cellular proteome. Dysregulation of protein phosphorylation signalling pathways is a major factor leading to different types of human disease, including cancers. Currently about 30% of all drug development programs across the pharmaceutical industry are focused on kinase inhibitors. So far only the skeleton of the body of protein phosphorylation pathways has been uncovered and the interaction maps between individual kinases, phosphatases and their substrates are far from clear. Despite major advances in phosporylation site discovery, utilizing enrichment strategies in combination with mass spectrometry, a major part of the phosphoproteome still remains undiscovered. In this proposal we seek to open a new window on the phosphoproteome by developing new methods for phosphopeptide enrichment and analysis by mass spectrometry, there by allowing the detection of previously undetected phosphorylation sites. As protein phosphorylation is one of the most important mechanisms for controlling cellular process, and kinase and phosphatase inhibitors are the focus of much pharmaceutical interest, the outcome of this study will benefit a wide range of investigators from both academic and pharmaceutical sectors. In the long term it will also benefit patients whose illness is a consequence of dysregulation of protein phosphorylation. In the short term the chemistry developed has the potential for commercial 'packaging' in a kit format.

可逆的蛋白质磷酸化已被证明是一种普遍的控制机制，几乎涉及细胞过程的所有方面。可逆性蛋白质磷酸化的重要性和复杂性可以从以下事实来评估：人类基因组中存在至少518种蛋白质激酶和100多种蛋白质磷酸酶，并且估计在给定的细胞蛋白质组中有三分之一的蛋白质被磷酸化。蛋白质磷酸化信号通路的失调是导致不同类型的人类疾病（包括癌症）的主要因素。目前，制药行业约30%的药物开发计划集中在激酶抑制剂上。到目前为止，蛋白质磷酸化途径的研究还只涉及到蛋白质磷酸化途径的基本框架，各个激酶、磷酸酶与底物之间的相互作用图谱还很不清楚。尽管在磷酸化位点发现方面取得了重大进展，利用富集策略与质谱法相结合，但磷酸化蛋白质组的主要部分仍然未被发现。在这个建议中，我们试图打开一个新的窗口，通过开发新的方法，磷酸肽富集和质谱分析的磷酸化蛋白质组，有允许以前未检测到的磷酸化位点的检测。由于蛋白质磷酸化是控制细胞过程的最重要机制之一，而激酶和磷酸酶抑制剂是许多药学兴趣的焦点，因此本研究的结果将使学术和药学领域的广泛研究人员受益。从长远来看，它也将使那些因蛋白质磷酸化失调而患病的患者受益。在短期内，开发的化学物质具有以试剂盒形式进行商业“包装”的潜力。

项目成果

Identification of LXR interacting proteins

LXR 相互作用蛋白的鉴定

• 作者: William Griffiths (Co-Author)

24S,25-Epoxycholesterol in mouse and rat brain.

• DOI: 10.1016/j.bbrc.2014.05.012
• 发表时间: 2014-06-27
• 期刊: BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
• 影响因子: 3.1
• 作者: Wang, Yuchen;Karu, Kersti;Meljon, Anna;Turton, John;Yau, Joyce L.;Seckl, Jonathan R.;Wang, Yuqin;Griffiths, William J.
• 通讯作者: Griffiths, William J.