Grant: Strategic Projects - Group

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 several kinases simultaneously. We will focus on protein kinases linked to breast cancer and develop an electrochemical microchip able to probe the activity of four key protein kinases. This electrochemical approach is at the cutting edge and has the potential to supersede existing technologies, and will undoubtedly find widespread application in cancer diagnostics and in monitoring patient responses to therapy. We foresee applications in the field of drug development through its amenability for automated high throughput screening of potential kinase inhibitor candidates. Given the role of protein kinases in tumorigenesis and their emergence as the largest potential family of «druggable» enzymes, this research is expected to have a direct impact on cancer diagnosis and treatment.