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Protein Kinase C-delta Inhibitor Peptide Formulation using Gold Nanoparticles
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Simple and Sensitive Method for Determination of Protein Kinase Activity Based on Surface Charge Change of

Fang Yi1, Xiangyi Huang1, Jicun Ren1

  • 1School of Chemistry and Chemical Engineering, State Key Laboratory of Metal Matrix Composites , Shanghai Jiao Tong University , 800 Dongchuan Road , Shanghai , 200240 , People's Republic of China.

Analytical Chemistry
|February 23, 2018
PubMed

Insights

This study introduces a novel, sensitive assay for detecting protein kinase activity using peptide-modified gold nanoparticles. This method offers a simple and cost-effective platform for drug discovery and fundamental research on kinases.

Area of Science:

  • Biochemistry
  • Nanotechnology
  • Molecular Biology

Background:

  • Protein tyrosine kinases are crucial in cell signaling and cancer development.
  • Existing kinase assays can be complex, costly, and lack sensitivity.
  • Developing simple, sensitive assays is vital for kinase research and drug discovery.

Purpose of the Study:

  • To present a novel, homogeneous assay for detecting protein kinase activity.
  • To enable the screening of kinase inhibitors using a cost-effective method.
  • To utilize gold nanoparticles (GNPs) as substrates for kinase detection.

Main Methods:

  • Peptide-modified gold nanoparticles (GNPs) were used as substrates for kinase detection.
  • Protein kinase activity was measured by changes in the surface charge (zeta potential) of GNPs upon phosphorylation.
  • Abelson murine leukemia viral oncogene (Abl) kinase served as a model system.

Main Results:

  • Phosphorylation of peptide-modified GNPs by Abl kinase significantly increased their surface negative charge.
  • The assay demonstrated a linear response to Abl kinase concentration within the range of 1–40 nM, with a detection limit of 1 nM.
  • The method accurately evaluated inhibitor efficiency, yielding IC50 values consistent with literature, and was successfully applied to cell lysates.

Conclusions:

  • This new method provides a simple, rapid, and highly sensitive approach for homogeneous detection of protein kinase activity.
  • The assay is suitable for screening kinase inhibitors and determining kinase activity in biological samples.
  • The developed platform holds promise for advancing kinase-related research and drug discovery efforts.

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