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

Nikhil Bhalla, Mirella Di Lorenzo, Giordano Pula, Pedro Estrela

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

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-α 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.

LanguageEnglish
Pages109-114
Number of pages6
JournalBiosensors and Bioelectronics
Volume54
DOIs
Publication statusPublished - 31 Oct 2014

Fingerprint

Phosphorylation
Drug Discovery
Protons
Phosphotransferases
Proteins
Phospho-Specific Antibodies
Throughput
Physiological Phenomena
Semiconductors
Myelin Basic Protein
Eukaryotic Cells
Post Translational Protein Processing
Antibodies
Oxides
Biomedical Research
Assays
Electrodes
Enzymes
Innovation
Hydrogen

Keywords

  • Drug discovery
  • PH sensors
  • Protein phosphorylation

Cite this

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Protein phosphorylation analysis based on proton release detection : Potential tools for drug discovery. / Bhalla, Nikhil; Di Lorenzo, Mirella; Pula, Giordano; Estrela, Pedro.

In: Biosensors and Bioelectronics, Vol. 54, 31.10.2014, p. 109-114.

Research output: Contribution to journalArticle

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