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Calmodulin-dependent Signaling

Calmodulin-dependent Signaling

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.
The Ca2+-CaM complex does not have enzymatic activity by itself. Instead, the complex binds downstream target proteins, including membrane proteins or enzymes,...

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...

Covalently Linked Protein Regulators

Covalently Linked Protein Regulators

Proteins can undergo many types of post-translational modifications, often in response to changes in their environment. These modifications play an important role in the function and stability of these proteins. Covalently linked molecules include functional groups, such as methyl, acetyl, and phosphate groups, and also small proteins, such as ubiquitin. There are around 200 different types of covalent regulators that have been identified.
These groups modify specific amino acids in a protein.

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...

Cooperative Allosteric Transitions

Cooperative Allosteric Transitions

Cooperative allosteric transitions can occur in multimeric proteins, where each subunit of the protein has its own ligand-binding site. When a ligand binds to any of these subunits, it triggers a conformational change that affects the binding sites in the other subunits; this can change the affinity of the other sites for their respective ligands. The ability of the protein to change the shape of its binding site is attributed to the presence of a mix of flexible and stable segments in the...

Feedback Regulation of Calcium Concentration

Feedback Regulation of Calcium Concentration

Calcium is an essential signaling molecule required for various cellular functions. Calcium pumps and ion channels on cell and organellar membranes, such as those on the endoplasmic reticulum (ER), regulate calcium concentrations inside the cell. They remain closed, keeping the cytosolic calcium levels low at a resting state.
Various transmembrane receptors, such as G protein-coupled receptors (GPCRs), elicit a response to extracellular signals by increasing cytosolic calcium. Activated GPCRs...

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Related Experiment Video

Updated: May 19, 2026

Pull-down of Calmodulin-binding Proteins

Pull-down of Calmodulin-binding Proteins

Published on: January 23, 2012

Modulating uranium binding affinity in engineered calmodulin EF-hand peptides: effect of phosphorylation.

Romain Pardoux¹, Sandrine Sauge-Merle, David Lemaire

¹CEA, DSV IBEB, Laboratoire des Interactions Protéine-Métal, Saint-Paul-lez-Durance, France.

|August 8, 2012

Summary

Phosphorylation significantly enhances uranyl binding affinity in proteins, particularly at physiological pH. This finding is crucial for understanding uranium toxicity and protein interactions.

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Area of Science:

Biochemistry
Toxicology
Protein Chemistry

Background:

Uranium toxicity necessitates understanding uranyl binding to proteins.
Characterizing uranyl affinity determinants in proteins is essential.

Purpose of the Study:

To analyze the contribution of a phosphoryl group to uranium binding affinity.
To investigate uranyl binding to the EF-hand motif of calmodulin.

Main Methods:

Engineered calmodulin domain 1 to impair metal binding at site 2.
Phosphorylated threonine at position 9 in vitro using protein kinase CK2.
Determined uranyl and calcium binding affinities via tyrosine fluorescence and competition assays.

Main Results:

Phosphorylation increased uranyl affinity by ~5-fold at pH 6 (Kd = 5±1 nM).
At pH 7, affinity increased significantly (Kd = 0.25±0.06 nM), with direct phosphoryl-uranyl interaction.
FTIR and UV–Vis spectroscopy confirmed phosphoryl group involvement in uranyl binding.

Conclusions:

The phosphoryl group is a key determinant of uranyl binding affinity in proteins.
This interaction is pH-dependent and significant at physiological pH.
Findings contribute to understanding uranium's biological interactions and toxicity.