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The Effect of Ca 2+ , Lobe-Specificity, and CaMKII on CaM Binding to Na V 1.1.
- Source :
-
International journal of molecular sciences [Int J Mol Sci] 2018 Aug 23; Vol. 19 (9). Date of Electronic Publication: 2018 Aug 23. - Publication Year :
- 2018
-
Abstract
- Calmodulin (CaM) is well known as an activator of calcium/calmodulin-dependent protein kinase II (CaMKII). Voltage-gated sodium channels (VGSCs) are basic signaling molecules in excitable cells and are crucial molecular targets for nervous system agents. However, the way in which Ca <superscript>2+</superscript> /CaM/CaMKII cascade modulates Na <subscript>V</subscript> 1.1 IQ (isoleucine and glutamine) domain of VGSCs remains obscure. In this study, the binding of CaM, its mutants at calcium binding sites (CaM <subscript>12</subscript> , CaM <subscript>34</subscript> , and CaM <subscript>1234</subscript> ), and truncated proteins (N-lobe and C-lobe) to Na <subscript>V</subscript> 1.1 IQ domain were detected by pull-down assay. Our data showed that the binding of Ca <superscript>2+</superscript> /CaM to the Na <subscript>V</subscript> 1.1 IQ was concentration-dependent. ApoCaM (Ca <superscript>2+</superscript> -free form of calmodulin) bound to Na <subscript>V</subscript> 1.1 IQ domain preferentially more than Ca <superscript>2+</superscript> /CaM. Additionally, the C-lobe of CaM was the predominant domain involved in apoCaM binding to Na <subscript>V</subscript> 1.1 IQ domain. By contrast, the N-lobe of CaM was predominant in the binding of Ca <superscript>2+</superscript> /CaM to Na <subscript>V</subscript> 1.1 IQ domain. Moreover, CaMKII-mediated phosphorylation increased the binding of Ca <superscript>2+</superscript> /CaM to Na <subscript>V</subscript> 1.1 IQ domain due to one or several phosphorylation sites in T1909, S1918, and T1934 of Na <subscript>V</subscript> 1.1 IQ domain. This study provides novel mechanisms for the modulation of Na <subscript>V</subscript> 1.1 by the Ca <superscript>2+</superscript> /CaM/CaMKII axis. For the first time, we uncover the effect of Ca <superscript>2+</superscript> , lobe-specificity and CaMKII on CaM binding to Na <subscript>V</subscript> 1.1.
- Subjects :
- Amino Acid Sequence
Binding Sites
Calcium metabolism
Calcium-Calmodulin-Dependent Protein Kinase Type 2 genetics
Calcium-Calmodulin-Dependent Protein Kinase Type 2 metabolism
Calmodulin genetics
Calmodulin metabolism
Cloning, Molecular
Crystallography, X-Ray
Escherichia coli genetics
Escherichia coli metabolism
Gene Expression
Genetic Vectors chemistry
Genetic Vectors metabolism
Glutathione Transferase chemistry
Glutathione Transferase genetics
Glutathione Transferase metabolism
HEK293 Cells
Humans
Kinetics
Molecular Docking Simulation
Mutation
NAV1.1 Voltage-Gated Sodium Channel genetics
NAV1.1 Voltage-Gated Sodium Channel metabolism
Phosphorylation
Protein Binding
Protein Conformation, alpha-Helical
Protein Conformation, beta-Strand
Protein Interaction Domains and Motifs
Recombinant Fusion Proteins chemistry
Recombinant Fusion Proteins genetics
Recombinant Fusion Proteins metabolism
Thermodynamics
Calcium chemistry
Calcium-Calmodulin-Dependent Protein Kinase Type 2 chemistry
Calmodulin chemistry
NAV1.1 Voltage-Gated Sodium Channel chemistry
Subjects
Details
- Language :
- English
- ISSN :
- 1422-0067
- Volume :
- 19
- Issue :
- 9
- Database :
- MEDLINE
- Journal :
- International journal of molecular sciences
- Publication Type :
- Academic Journal
- Accession number :
- 30142967
- Full Text :
- https://doi.org/10.3390/ijms19092495