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Protein Kinases and Phosphatases

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...

Phosphorylation

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...

Phosphorylation

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: Jul 7, 2026

Oligopeptide Competition Assay for Phosphorylation Site Determination

Oligopeptide Competition Assay for Phosphorylation Site Determination

Published on: May 18, 2017

Immunological detection of phosphorylation.

Z Yao¹, R Seger

¹The Weizmann Institute of Science, Rehovot, Israel.

Current Protocols in Cell Biology

|January 30, 2008

Summary

Detecting protein phosphorylation is crucial for understanding protein function. This study details methods using antibodies to identify protein phosphorylation and kinase activity, with immunoblotting being a convenient option.

Area of Science:

Biochemistry
Molecular Biology
Cell Signaling

Background:

Protein phosphorylation, the addition of phosphates to serine, threonine, or tyrosine residues, is a key regulatory mechanism controlling protein function.
Understanding protein phosphorylation is vital for deciphering cellular signaling pathways and disease mechanisms.

Purpose of the Study:

To present reliable protocols for detecting protein phosphorylation and protein kinase activity.
To evaluate different immunological methods for identifying phosphorylated proteins.

Main Methods:

Immunoblotting using specific anti-phosphoamino acid (PAA) or anti-phosphoprotein antibodies.
Immunoprecipitation followed by immunoblot analysis.
Immunofluorescent staining.

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

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

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Last Updated: Jul 7, 2026

Oligopeptide Competition Assay for Phosphorylation Site Determination

Oligopeptide Competition Assay for Phosphorylation Site Determination

Published on: May 18, 2017

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

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

Published on: June 6, 2022

Main Results:

Immunoblotting provides a convenient and effective method for detecting protein phosphorylation.
While more complex, immunoprecipitation and immunofluorescence also successfully detect protein phosphorylation.
All presented methods are versatile, working with various antibodies and phosphorylated proteins.

Conclusions:

Specific antibodies are valuable tools for detecting protein phosphorylation and kinase activity.
Immunoblotting is a highly recommended method for its convenience and effectiveness.
The choice of method depends on experimental needs, balancing complexity with desired outcomes.