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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.
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Protein Kinases and Phosphatases

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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...

Protein Kinases and Phosphatases

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cAMP-dependent Protein Kinase Pathways

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Cyclic Adenosine Monophosphate (cAMP) is an essential second messenger that activates protein kinase A (PKA) and regulates various biological processes. A single epinephrine molecule binds to GPCR and activates several heterotrimeric G proteins, each stimulating multiple adenylyl cyclase, amplifying the signal, and synthesizing large numbers of cAMP molecules. Small changes in cAMP concentration affect PKA activity. The binding of four cAMP molecules induces a conformational change in PKA,...

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Calmodulin (CaM) is a calcium-binding protein in eukaryotes that controls various calcium-regulated cellular processes. It has four calcium-binding sites that bind calcium to form the calcium-calmodulin ( Ca2+-CaM) complex. GPCR stimulation increases the calcium levels in the cells that bind to CaM and induces a conformational change.
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Related Experiment Video

Updated: Jul 27, 2026

Detecting Anastasis In Vivo by CaspaseTracker Biosensor

Detecting Anastasis In Vivo by CaspaseTracker Biosensor

Published on: February 1, 2018

Imaging protein kinase Calpha activation in cells.

T Ng¹, A Squire, G Hansra

¹Protein Phosphorylation Laboratory and Cell Biophysics Laboratory, Imperial Cancer Research Fund (ICRF), 44 Lincoln's Inn Fields, London, WC2A 3PX, UK.

Science (New York, N.Y.)

|March 26, 1999

Summary

This study identifies a new way to track protein kinase Calpha (PKCalpha) activation using fluorescence resonance energy transfer (FRET) and microscopy. This method visualizes enzyme activity in real-time within cells and tissues.

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Area of Science:

Biophysics
Cell Biology
Biochemistry

Background:

Fluorescence resonance energy transfer (FRET) and fluorescence lifetime imaging microscopy (FLIM) offer powerful tools for studying protein function.
Spatially resolved FRET/FLIM can monitor protein activity in live and fixed biological samples.

Purpose of the Study:

To develop and validate a FRET/FLIM-based method for dynamically imaging protein kinase Calpha (PKCalpha) activation.
To demonstrate the utility of this method in live cells, fixed cells, and pathological tissues.

Main Methods:

Utilized FRET measured by FLIM to detect PKCalpha activation.
Employed fluorescently tagged phosphorylation site-specific antibodies to identify activated PKCalpha.
Applied the technique to live and fixed cultured cells, as well as pathological samples.

Main Results:

Successfully identified and exploited a dynamic marker for PKCalpha activation.
Enabled real-time imaging of PKCalpha activation in cellular environments.
Demonstrated applicability to both experimental cell cultures and clinical pathological samples.

Conclusions:

Spatially resolved FRET/FLIM is a viable method for tracing protein catalytic activity and functional states.
This approach provides a dynamic readout of PKCalpha activation.
The method has broad potential for biological and clinical research.