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Updated: May 29, 2026
08:07
Probing High-density Functional Protein Microarrays to Detect Protein-protein Interactions
Published on: August 2, 2015
Probing calmodulin protein-protein interactions using high-content protein arrays
David J O'Connell1, Mikael Bauer, Sara Linse
1Conway Institute of Biomolecular & Biomedical Research, University College Dublin, Dublin, Ireland.
Methods in Molecular Biology (Clifton, N.J.)
|September 9, 2011
Summary
Calcium ion (Ca2+) is vital for cell signaling, regulated by proteins like calmodulin. Researchers discovered calmodulin binds strongly to STIM1 and STIM2, key proteins in calcium entry into cells.
Area of Science:
- Cellular Biology
- Biochemistry
- Molecular Biology
Background:
- Calcium ion (Ca2+) acts as a critical second messenger regulating diverse cellular functions.
- Intracellular Ca2+-binding proteins, notably calmodulin, mediate Ca2+ signals.
- Calmodulin is essential for cellular responses to stimuli that increase cytosolic Ca2+ concentration.
Purpose of the Study:
- To identify novel protein-protein interactions involving calmodulin.
- To investigate calmodulin's role in cellular calcium regulation pathways.
Main Methods:
- Utilized a high-content recombinant human protein array.
- Probed the array with fluorophore-labeled calmodulin in the presence of Ca2+.
- Screened over 10,000 unique human proteins from a human brain cDNA library.
Main Results:
- Identified a high-affinity interaction between calmodulin and STIM1.
- Identified a high-affinity interaction between calmodulin and STIM2.
- STIM1 and STIM2 are single-pass transmembrane proteins localized to the endoplasmic reticulum.
Conclusions:
- Calmodulin directly interacts with STIM1 and STIM2.
- This interaction is significant for the translocation of STIM1/STIM2 to the plasma membrane.
- The findings shed light on the regulation of store-operated calcium entry.
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