Electrochemical biosensors based on peptide-kinase interactions at the kinase docking site

Pralhad Namdev Joshi, Evgeniy Mervinetsky, Ohad Solomon, Yu Ju Chen, Shlomo Yitzchaik, Assaf Friedler

Research output: Contribution to journalArticlepeer-review

Abstract

Kinases are important cancer biomarkers and are conventionally detected based on their catalytic activity. Kinases regulate cellular activities by phosphorylation of motif-specific multiple substrate proteins, resulting in a lack of selectivity of activity-based kinase biosensors. We present an alternative approach of sensing kinases based on the interactions of their allosteric docking sites with a specific partner protein. The new approach was demonstrated for the ERK2 kinase and its substrate ELK-1. A peptide derived from ELK-1 was bound to a gold electrode and ERK2 sensing was performed by electrochemical impedance spectroscopy. We performed a detailed analysis of the interaction between the ELK-1 peptide and the kinase on gold surfaces. Atomic force microscopy, variable angle spectroscopic ellipsometry, X-ray Photoelectron Spectroscopy, and polarization modulation IR reflection-absorption spectroscopy analysis of the gold surface revealed the adsorbed layer of the ERK2 on the peptide monolayer. The sensors showed a high level of target selectivity for ERK2 compared to the p38γ kinase and BSA. ERK2 was detected in its cellular concentration range, 0.5–2.0 μM, and the limit of detection was calculated to be 0.35 μM. Using the flexibility of peptide design, our method is generic for developing sensitive and substrate-specific biosensors and other disease-related enzymes based on their interactions.

Original languageAmerican English
Article number114177
Pages (from-to)114177
JournalBiosensors and Bioelectronics
Volume207
DOIs
StatePublished - 1 Jul 2022

Keywords

  • Amino Acid Sequence
  • Biomarkers
  • Biosensing Techniques
  • Electrochemical biosensors
  • Gold
  • Kinases
  • Peptide self-assembly
  • Peptide-protein interactions
  • Peptides/chemistry
  • Phosphorylation
  • Protein-protein interactions (PPI)

All Science Journal Classification (ASJC) codes

  • Biophysics
  • Biotechnology
  • Biomedical Engineering
  • Electrochemistry

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