Electrochemical method for monitoring kinase activity

电化学方法监测激酶活性

• 批准号: 365192-2008
• 负责人: Kraatz, HeinzBernhard
• 金额: $ 3.62万
• 依托单位: University of Toronto
• 依托单位国家: 加拿大
• 项目类别: Strategic Projects - Group
• 财政年份: 2011
• 资助国家: 加拿大
• 起止时间: 2011-01-01 至 2012-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=483403
• 关键词: Electrochemical; method; monitoring; kinase; activity

Kinases are enzymes that are involved in regulating key cell functions. The reaction catalyzed by kinases is the transfer of a phosphate group from adenosine triphosphate to a hydroxy amino acid. Their activity is carefully controlled and uncontrolled activation of these enzymes has been linked to various forms of cancer. In our proposal, we make use of this fundamental reaction and propose a method for the rapid monitoring and profiling the kinase activity in cell and tissue extracts using. This screening method exploits our recent discovery of the protein kinase catalyzed transfer of a redox label from a novel redox-active analogue of adenosine triphosphate. This approach will enable the quantification of the kinase activity by electrochemical methods. In our approach, we make use of an electrochemical peptide array that has individually addressable gold pads, each having a peptide specific for a particular protein kinase, allowing us to monitor the activity of

激酶是参与调节关键细胞功能的酶。激酶催化的反应是磷酸基团从三磷酸腺苷转移到羟基氨基酸。它们的活性受到仔细控制，这些酶的不受控制的激活与各种形式的癌症有关。在我们的建议中，我们利用这一基本反应，并提出了一种方法，用于快速监测和分析细胞和组织提取物中的激酶活性。这种筛选方法利用了我们最近发现的蛋白激酶催化转移的氧化还原标记从一个新的氧化还原活性类似物的三磷酸腺苷。这种方法将能够通过电化学方法定量激酶活性。在我们的方法中，我们利用电化学肽阵列，其具有可单独寻址的金垫，每个金垫具有对特定蛋白激酶特异的肽，使我们能够监测蛋白激酶的活性。