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Related Concept Videos

Protein Kinases and Phosphatases02:54

Protein Kinases and Phosphatases

Proteins undergo chemical modifications that trigger changes in the charge, structure, and conformation of the proteins. Phosphorylation, acetylation, glycosylation, nitrosylation, ubiquitination, lipidation, methylation, and proteolysis are various protein modifications that regulate protein activity. Such modifications are usually enzyme-driven.
Protein kinases
Many proteins in the cell are regulated by phosphorylation, the addition of a phosphate group. A family of enzymes called kinases...
Phosphorylation01:02

Phosphorylation

The addition or removal of phosphate groups from proteins is the most common chemical modification that regulates cellular processes. These modifications can affect the structure, activity, stability, and localization of proteins within cells as well as their interactions with other proteins.
During phosphorylation, protein kinases transfer the terminal phosphate group of ATP to specific amino acid side chains of substrate proteins. Serine, threonine, and tyrosine are the most commonly...

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Related Experiment Video

Updated: May 26, 2026

Oligopeptide Competition Assay for Phosphorylation Site Determination
09:16

Oligopeptide Competition Assay for Phosphorylation Site Determination

Published on: May 18, 2017

Electrochemical strategy for sensing protein phosphorylation.

Peng Miao1, Limin Ning, Xiaoxi Li

  • 1Laboratory of Biosensing Technology, School of Life Sciences, Shanghai University, Shanghai, P.R. China.

Bioconjugate Chemistry
|December 14, 2011
PubMed
Summary

This study introduces a new electrochemical method to detect protein phosphorylation and kinase activity. It uses zirconium ions and rolling circle amplification for sensitive enzyme assays.

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An Optimized Single-Molecule Pull-Down Assay for Quantification of Protein Phosphorylation
07:45

An Optimized Single-Molecule Pull-Down Assay for Quantification of Protein Phosphorylation

Published on: June 6, 2022

Related Experiment Videos

Last Updated: May 26, 2026

Oligopeptide Competition Assay for Phosphorylation Site Determination
09:16

Oligopeptide Competition Assay for Phosphorylation Site Determination

Published on: May 18, 2017

An Optimized Single-Molecule Pull-Down Assay for Quantification of Protein Phosphorylation
07:45

An Optimized Single-Molecule Pull-Down Assay for Quantification of Protein Phosphorylation

Published on: June 6, 2022

Area of Science:

  • Biochemistry
  • Electrochemistry
  • Molecular Biology

Background:

  • Protein phosphorylation is a critical cellular signaling mechanism.
  • Assaying protein kinase activity is essential for understanding cellular processes and disease.
  • Existing methods for monitoring phosphorylation and kinase activity have limitations.

Purpose of the Study:

  • To develop a novel, sensitive electrochemical method for monitoring protein phosphorylation.
  • To establish a robust assay for protein kinase activity using electrochemical detection.
  • To leverage signal amplification strategies for enhanced detection sensitivity.

Main Methods:

  • Immobilization of substrate peptide on a gold electrode.
  • Enzymatic phosphorylation by protein kinase A.
  • Zirconium (Zr4+) mediated linkage of phosphorylated peptide to a DNA primer probe.
  • Rolling circle amplification (RCA) initiated by the primer on the electrode surface.
  • Electrochemical detection using [Ru(NH3)6]3+ as a redox indicator.

Main Results:

  • The method successfully detected protein kinase A activity.
  • The electrochemical signal correlated with the extent of phosphorylation.
  • A low detection limit of 0.5 unit/mL was achieved for protein kinase A.
  • The method demonstrated high sensitivity and specificity.

Conclusions:

  • The developed electrochemical method offers a sensitive platform for monitoring protein phosphorylation.
  • This technique provides a promising approach for assaying protein kinase activity.
  • The method holds potential for future applications in biological and medical research.