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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...

Phosphoinositides and PIPs

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Phosphoinositides are a group of phospholipids containing a glycerol backbone with two fatty acid chains and a phosphate attached to a myoinositol sugar ring. The inositol head group extends into the cytoplasm, where it is modified by adding phosphate groups to form phosphatidylinositol phosphates or PIPs.
Different phosphoinositides are synthesized and recruited on the cytosolic face of the plasma membrane. The localization of specific phosphoinositides concentrated in separate membrane...

Protein-protein Interfaces

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Many proteins form complexes to carry out their functions, making protein-protein interactions (PPIs) essential for an organism's survival. Most PPIs are stabilized by numerous weak noncovalent chemical forces. The physical shape of the interfaces determines the way two proteins interact. Many globular proteins have closely-matching shapes on their surfaces, which form a large number of weak bonds. Additionally, many PPIs occur between two helices or between a surface cleft and a...

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Allosteric proteins have more than one ligand binding site; the binding of a ligand to any of these sites influences the binding of ligands to the other sites. When a protein is allosteric, its binding sites are called coupled or linked. In the case of enzymes, the site that binds to the substrate is known as the active site and the other site is known as the regulatory site. When a ligand binds to the regulatory site, this leads to conformational changes in the protein that can influence...

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Related Experiment Video

Updated: Feb 23, 2026

Identification of Post-translational Modifications of Plant Protein Complexes

Identification of Post-translational Modifications of Plant Protein Complexes

Published on: February 22, 2014

Phosphate Transfer in Activated Protein Complexes Reveals Interaction Sites.

Sem Tamara¹, Richard A Scheltema¹, Albert J R Heck¹

¹Biomolecular Mass Spectrometry and Proteomics, Bijvoet Center for Biomolecular Research and Utrecht Institute for Pharmaceutical Sciences and Netherlands Proteomics Centre, Utrecht University, Padualaan 8, 3584 CH, Utrecht, The Netherlands.

Angewandte Chemie (International Ed. in English)

|September 5, 2017

Summary

This study reveals non-enzymatic phosphate transfer between interacting proteins in the gas phase. This unexpected mechanism precisely defines protein binding sites and is retained during phase transfer.

Keywords:

Pin1 native MS phosphorylation protein-protein interactions proteomics

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Identification of Post-translational Modifications of Plant Protein Complexes

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Oligopeptide Competition Assay for Phosphorylation Site Determination

Oligopeptide Competition Assay for Phosphorylation Site Determination

Published on: May 18, 2017

Area of Science:

Biochemistry
Chemical Physics
Molecular Interactions

Background:

Phosphorylation is a key post-translational modification regulating protein interactions.
Understanding protein-binding interfaces is crucial for deciphering cellular signaling pathways.

Purpose of the Study:

To investigate non-enzymatic phosphate group transfer between interacting biomolecules.
To explore the gas-phase behavior of protein-ligand interactions and binding site definition.

Main Methods:

Utilized gas-phase activation techniques to induce non-enzymatic reactions.
Analyzed phosphate transfer dynamics at the molecular interaction interface.

Main Results:

Observed efficient, non-enzymatic transfer of phosphate groups between binding partners in the gas phase.
Demonstrated that phosphate transfer is highly specific to proximity within the binding interface.
Showcased the ability of this transfer to precisely define protein-ligand binding sites.

Conclusions:

Non-enzymatic phosphate transfer can occur efficiently in the gas phase between high-affinity interacting proteins.
This phenomenon offers a novel method for high-precision mapping of protein-protein binding sites.
The observed binding site definition is robust, persisting through phase transfer from solution to gas phase.