Protein phosphorylation analysis based on proton release detection:potential tools for drug discovery


Bhalla, N., Di Lorenzo, M., Pula, G. and Estrela, P., 2014. Protein phosphorylation analysis based on proton release detection:potential tools for drug discovery. Biosensors and Bioelectronics, 54, pp. 109-114.

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    Phosphorylation is the most important post-translational modification of proteins in eukaryotic cells and it is catalysed by enzymes called kinases. The balance between protein phosphorylation and dephosphorylation is critical for the regulation of physiological processes and its unbalance is the cause of several diseases. Conventional assays used to analyse the kinase activity are limited as they rely heavily on phospho-specific antibodies and radioactive tags. This makes their use impractical for high throughput drug discovery platforms. We have developed two versatile methods to detect the release of protons (H+) associated with the protein phosphorylation catalysed by kinases. The first approach is based on the pH-sensitive response of oxide-semiconductor interfaces and the second method detects the pH changes in phosphorylation reaction using a commercial micro pH electrode. The proposed methods successfully detected phosphorylation of myelin basic protein by PKC- kinase. These techniques can be readily adopted for multiplexed arrays and high throughput analysis of kinase activity, which will represent an important innovation in biomedical research and drug discovery.


    Item Type Articles
    CreatorsBhalla, N., Di Lorenzo, M., Pula, G. and Estrela, P.
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    Uncontrolled Keywordsprotein phosphorylation,ph sensors,drug discovery
    DepartmentsFaculty of Engineering & Design > Electronic & Electrical Engineering
    Faculty of Engineering & Design > Chemical Engineering
    Faculty of Science > Pharmacy & Pharmacology
    Research CentresCentre for Sustainable Chemical Technologies
    Centre for Regenerative Medicine
    Centre for Advanced Sensor Technologies (CAST)
    Publisher StatementBhalla_et_al_Biosensors_Bioelectronics_2013_54.pdf: NOTICE: this is the author’s version of a work that was accepted for publication in Biosensors and Bioelectronics. Changes resulting from the publishing process, such as peer review, editing, corrections, structural formatting, and other quality control mechanisms may not be reflected in this document. Changes may have been made to this work since it was submitted for publication. A definitive version was subsequently published in Biosensors and Bioelectronics, vol 54, 2014, DOI 10.1016/j.bios.2013.10.037
    ID Code37752


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