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Assessment of Resistance to Tyrosine Kinase Inhibitors by an Interrogation of Signal Transduction Pathways by Antibody Arrays
Published on: September 19, 2018
Detection of aberrant protein phosphorylation in cancer using direct gold-protein affinity interactions
Mostak Ahmed1, Laura G Carrascosa1, Abu Ali Ibn Sina1
1Centre for Personalized Nanomedicine, Australian Institute for Bioengineering and Nanotechnology (AIBN), The University of Queensland, Corner College and Cooper Roads (Bldg 75), Brisbane QLD 4072, Australia.
Abstract:
Protein phosphorylation is one of the most prominent post-translational mechanisms for protein regulation, which is frequently impaired in cancer. Through the covalent addition of phosphate groups to certain amino-acids, the interactions of former residues with nearby amino-acids are drastically altered, resulting in major changes of protein conformation that impacts its biological function. Herein, we report that these conformational changes can also disturb the protein's ability to interact with and adsorb onto bare gold surfaces. We exploited this feature to develop a simple electrochemical method for detecting the aberrant phosphorylation of EGFR protein in several lung cancer cell lines. This method, which required as low as 10ng/µL (i.e., 50ng) of purified EGFR protein, also enabled monitoring cell sensitivity to tyrosine kinase inhibitors (TKI) - a common drug used for restoring the function of aberrantly phosphorylated proteins in lung cancer. The reported strategy based on direct gold-protein affinity interactions avoids the conventional paradigm of requiring a phospho-specific antibody for detection and could be a potential alternative of widely used mass spectrometry.
Insights
This study introduces a novel electrochemical method to detect aberrant protein phosphorylation in cancer by observing changes in gold surface interactions. This approach offers a simpler alternative for monitoring cancer cell sensitivity to drugs like tyrosine kinase inhibitors (TKIs).
Area of Science:
- Biochemistry
- Oncology
- Electrochemistry
Background:
- Protein phosphorylation is a key post-translational modification regulating protein function, often dysregulated in cancer.
- Altered protein conformation due to phosphorylation can affect interactions with surfaces, including gold.
- Aberrant phosphorylation of Epidermal Growth Factor Receptor (EGFR) is implicated in lung cancer progression.
Purpose of the Study:
- To develop a simple electrochemical method for detecting aberrant EGFR phosphorylation in lung cancer.
- To assess the utility of this method in monitoring cancer cell sensitivity to tyrosine kinase inhibitors (TKIs).
Main Methods:
- Exploiting altered protein-gold surface adsorption due to phosphorylation-induced conformational changes.
- Developing an electrochemical detection strategy for EGFR phosphorylation.
- Utilizing purified EGFR protein at low concentrations (10ng/µL).
Main Results:
- Demonstrated a novel electrochemical method for detecting aberrant EGFR phosphorylation in lung cancer cell lines.
- Successfully monitored cancer cell sensitivity to TKIs using the developed method.
- The method requires minimal purified EGFR protein and avoids the need for phospho-specific antibodies.
Conclusions:
- The developed electrochemical strategy offers a direct gold-protein affinity-based detection of aberrant protein phosphorylation.
- This method provides a potential alternative to mass spectrometry and antibody-dependent assays for cancer biomarker detection.
- The approach facilitates monitoring of therapeutic responses to TKIs in lung cancer.
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