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Book of Abstracts - Ruhr-Universität Bochum

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P-62<br />

ISBOMC `10 5.7 – 9.7. 2010 <strong>Ruhr</strong>-<strong>Universität</strong> <strong>Bochum</strong><br />

Electrochemical Detection <strong>of</strong> Protein Kinases and Inhibitor Screening by Using<br />

Ferrocene-ATP Bioconjugate<br />

Sanela Martić, Mahmoud Labib and Heinz-Bernhard Kraatz*<br />

The University <strong>of</strong> Western Ontario, Faculty <strong>of</strong> Science, Department <strong>of</strong> Chemistry, 1151 Richmond<br />

Street, N6A 5B7, London, Canada.<br />

E-mail: smartic@uwo.ca<br />

Protein kinase plays a critical role in the cellular growth, signalling and function and its malfunction<br />

has been linked to a number <strong>of</strong> diseases, such as cancer. 1 For diagnostic and drug screening purposes<br />

detecting, monitoring and quantifying kinase activity is critical. Recently, an electrochemical<br />

biosensor was developed and was used in our laboratory for measuring casein kinase 2 (CK2) and<br />

protein kinase C (PKC) activity on a Au surface by means <strong>of</strong> the electroactive adenosine-5’-[γferrocene]<br />

triphosphate conjugate (Fc-C6-ATP), as a co-substrate for the protein kinase. 2 As an<br />

extension <strong>of</strong> this work, we have investigated following kinases: sarcoma-related kinase (Src), cyclindependent<br />

kinase (CDK2) and extracellular signal–regulated kinase (Erk1), all <strong>of</strong> which are directly<br />

involved in the cell cycle. The electrochemical detection <strong>of</strong> kinase activity and drug target screening<br />

was performed in addition to the surface characterization <strong>of</strong> phosphorylated assays by MALDI-TOF<br />

MS, XPS and TOF-SIMS techniques. A pro<strong>of</strong>-<strong>of</strong>-concept study was performed using the newly<br />

developed multiplex chip technology which allows for monitoring and quantifying multiple kinase<br />

activities for the first time. The optimized electrochemical multiplex assay was also used for<br />

identification <strong>of</strong> novel protein kinase inhibitors.<br />

References<br />

Erk1<br />

CDK2<br />

control<br />

Src<br />

120<br />

Fe<br />

O<br />

N<br />

H<br />

PO 3 Fc<br />

6<br />

N<br />

H<br />

ATP<br />

No electrochemical signal !<br />

Electrochemical signal<br />

after phosphorylation!<br />

1. (a) G. Manning, D. B. Whyte, R. Martinez, T. Hunter, S. Sudarsanam, Science, 2002, 298, 1912-<br />

1934. (b) J. S. Sebolt-Leopold, J. M. English, Nature, 2006, 441, 457-462.<br />

2. (a) H. Song, K. Kerman, H. B. Kraatz, Chem. Commun. 2008, 502-504. (b) K. Kerman, H. Song, J.<br />

D. Duncan, D. W. Litchfield, H. B. Kraatz, Anal. Chem. 2008, 80, 9395-9401.

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