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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...

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...

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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

Detection of phosphorylation by enzymatic techniques.

¹Duke University Medical Center, Durham, North Carolina, USA.

Current Protocols in Molecular Biology

|February 12, 2008

Summary

This study details methods for identifying protein phosphorylation, a key regulator in cells. Techniques involve using phosphatases to detect and characterize covalently bound phosphate in proteins.

Area of Science:

Biochemistry
Molecular Biology
Cellular Regulation

Background:

Reversible protein phosphorylation is a critical regulatory mechanism in both plant and animal cells.
Detecting covalently bound phosphate in proteins is essential for understanding physiological processes.
Existing techniques require specific methods to identify and characterize phosphoproteins.

Purpose of the Study:

To provide protocols for identifying and characterizing protein phosphorylation.
To demonstrate the functional significance of protein dephosphorylation using specific phosphatases.
To establish methods for distinguishing between different types of phosphorylated residues.

Main Methods:

Utilizing nonspecific acid or alkaline phosphatases to assess dephosphorylation effects.

More Related Videos

Characterization at the Molecular Level using Robust Biochemical Approaches of a New Kinase Protein

Characterization at the Molecular Level using Robust Biochemical Approaches of a New Kinase Protein

Published on: June 30, 2019

Related Experiment Videos

Last Updated: Jul 7, 2026

Oligopeptide Competition Assay for Phosphorylation Site Determination

Oligopeptide Competition Assay for Phosphorylation Site Determination

Published on: May 18, 2017

Characterization at the Molecular Level using Robust Biochemical Approaches of a New Kinase Protein

Characterization at the Molecular Level using Robust Biochemical Approaches of a New Kinase Protein

Published on: June 30, 2019

Employing selective protein phosphatases (hydrolyzing phosphoserine, phosphothreonine, or phosphotyrosine residues).

Describing protocols for digesting phosphoproteins with specific serine/threonine and tyrosine phosphatases.

Including a support protocol for identifying radiolabel as 32Pi.

Main Results:

Demonstrated the utility of nonspecific phosphatases in initial functional analysis.
Showcased the ability of selective phosphatases to identify specific phosphorylation sites.
Provided a method for confirming the radiolabel used in experiments.

Conclusions:

The described protocols offer a comprehensive approach to studying protein phosphorylation.
These methods enable the identification of functionally important covalent modifications.
The techniques are applicable to both plant and animal cell systems for phosphoprotein analysis.