Back to Search
Start Over
A general strategy to characterize calmodulin-calcium complexes involved in CaM-target recognition: DAPK and EGFR calmodulin binding domains interact with different calmodulin-calcium complexes.
- Source :
-
Biochimica et biophysica acta [Biochim Biophys Acta] 2011 May; Vol. 1813 (5), pp. 1059-67. Date of Electronic Publication: 2010 Nov 30. - Publication Year :
- 2011
-
Abstract
- Calmodulin (CaM) is a ubiquitous Ca(2+) sensor regulating many biochemical processes in eukaryotic cells. Its interaction with a great variety of different target proteins has led to the fundamental question of its mechanism of action. CaM exhibits four "EF hand" type Ca(2+) binding sites. One way to explain CaM functioning is to consider that the protein interacts differently with its target proteins depending on the number of Ca(2+) ions bound to it. To test this hypothesis, the binding properties of three entities known to interact with CaM (a fluorescent probe and two peptide analogs to the CaM binding sites of death associated protein kinase (DAPK) and of EGFR) were investigated using a quantitative approach based on fluorescence polarization (FP). Probe and peptide interactions with CaM were studied using a titration matrix in which both CaM and calcium concentrations were varied. Experiments were performed with SynCaM, a hybrid CaM able to activate CaM dependent enzymes from mammalian and plant cells. Results show that the interaction between CaM and its targets is regulated by the number of calcium ions bound to the protein, namely one for the DAPK peptide, two for the probe and four for the EGFR peptide. The approach used provides a new tool to elaborate a typology of CaM-targets, based on their recognition by the various CaM-Ca(n) (n=0-4) complexes. This article is part of a Special Issue entitled: 11th European Symposium on Calcium.<br /> (2010 Elsevier B.V. All rights reserved.)
- Subjects :
- Amino Acid Sequence
Death-Associated Protein Kinases
Fluorescent Dyes metabolism
Kinetics
Molecular Sequence Data
Mutant Proteins chemistry
Mutant Proteins metabolism
Peptides chemistry
Peptides metabolism
Protein Binding
Protein Structure, Tertiary
Recombinant Proteins chemistry
Recombinant Proteins metabolism
Static Electricity
Thermodynamics
Apoptosis Regulatory Proteins chemistry
Apoptosis Regulatory Proteins metabolism
Biochemistry methods
Calcium metabolism
Calcium-Calmodulin-Dependent Protein Kinases chemistry
Calcium-Calmodulin-Dependent Protein Kinases metabolism
Calmodulin metabolism
ErbB Receptors chemistry
ErbB Receptors metabolism
Subjects
Details
- Language :
- English
- ISSN :
- 0006-3002
- Volume :
- 1813
- Issue :
- 5
- Database :
- MEDLINE
- Journal :
- Biochimica et biophysica acta
- Publication Type :
- Academic Journal
- Accession number :
- 21115073
- Full Text :
- https://doi.org/10.1016/j.bbamcr.2010.11.004