BMRB Entry 19050
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PDB ID:
Entry in NMR Restraints Grid
Validation report in NRG-CING
Chem Shift validation: AVS_full, SPARTA
BMRB Entry DOI: doi:10.13018/BMR19050
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Title: Structure of the C-domain of Calcium-saturated Calmodulin bound to the IQ motif of NaV1.2 PubMed: 28343066
Deposition date: 2013-02-21 Original release date: 2014-07-21
Authors: Fowler, C.; Feldkamp, Michael; Yu, Liping; Shea, Madeline
Citation: Hovey, Liam; Fowler, C; Mahling, Ryan; Lin, Zesen; Miller, Mark; Marx, Dagan; Yoder, Jesse; Kim, Elaine; Tefft, Kristin; Waite, Brett; Feldkamp, Michael; Yu, Liping; Shea, Madeline. "Calcium triggers reversal of calmodulin on nested anti-parallel sites in the IQ motif of the neuronal voltage-dependent sodium channel NaV1.2." Biophys. Chem. 224, 1-19 (2017).
Assembly members:
C-domain_of_Calmodulin_(residues_76-148), polymer, 73 residues, 8416.369 Da.
IQ_motif_peptide_of_NaV1-2_(residues_1901-1927), polymer, 27 residues, 3370.125 Da.
CALCIUM ION, non-polymer, 40.078 Da.
Natural source: Common Name: Ciliates Taxonomy ID: 5888 Superkingdom: Eukaryota Kingdom: not available Genus/species: Paramecium tetraurelia
Experimental source: Production method: recombinant technology Host organism: Escherichia coli
Entity Sequences (FASTA):
C-domain_of_Calmodulin_(residues_76-148): MKEQDSEEELIEAFKVFDRD
GNGLISAAELRHVMTNLGEK
LTDDEVDEMIREADIDGDGH
INYEEFVRMMVSK
IQ_motif_peptide_of_NaV1-2_(residues_1901-1927): KRKQEEVSAIVIQRAYRRYL
LKQKVKK
- assigned_chemical_shifts
| Data type | Count |
| 13C chemical shifts | 314 |
| 15N chemical shifts | 78 |
| 1H chemical shifts | 606 |
Additional metadata:
Assembly:
| Entity Assembly ID | Entity Name | Entity ID |
|---|---|---|
| 1 | C-domain of Calmodulin | 1 |
| 2 | IQ motif peptide of NaV1.2 | 2 |
| 3 | CALCIUM ION_1 | 3 |
| 4 | CALCIUM ION_2 | 3 |
Entities:
Entity 1, C-domain of Calmodulin 73 residues - 8416.369 Da.
| 1 | MET | LYS | GLU | GLN | ASP | SER | GLU | GLU | GLU | LEU | ||||
| 2 | ILE | GLU | ALA | PHE | LYS | VAL | PHE | ASP | ARG | ASP | ||||
| 3 | GLY | ASN | GLY | LEU | ILE | SER | ALA | ALA | GLU | LEU | ||||
| 4 | ARG | HIS | VAL | MET | THR | ASN | LEU | GLY | GLU | LYS | ||||
| 5 | LEU | THR | ASP | ASP | GLU | VAL | ASP | GLU | MET | ILE | ||||
| 6 | ARG | GLU | ALA | ASP | ILE | ASP | GLY | ASP | GLY | HIS | ||||
| 7 | ILE | ASN | TYR | GLU | GLU | PHE | VAL | ARG | MET | MET | ||||
| 8 | VAL | SER | LYS |
Entity 2, IQ motif peptide of NaV1.2 27 residues - 3370.125 Da.
peptide containing the IQ motif of the voltage dependent sodium channel NaV1.2
| 1 | LYS | ARG | LYS | GLN | GLU | GLU | VAL | SER | ALA | ILE | ||||
| 2 | VAL | ILE | GLN | ARG | ALA | TYR | ARG | ARG | TYR | LEU | ||||
| 3 | LEU | LYS | GLN | LYS | VAL | LYS | LYS |
Entity 3, CALCIUM ION_1 - Ca - 40.078 Da.
| 1 | CA |
Samples:
sample_1: C-domain of Calmodulin, [U-100% 13C; U-100% 15N], 1.5 mM; IQ motif peptide of NaV1.2 1.5 mM; CALCIUM ION 3.3 mM; imidazole, [U-2H], 10 mM; potassium chloride 100 mM; sodium azide 0.01%; EDTA, [U-2H], 50 uM; H2O 95%; D2O 5%
sample_2: C-domain of Calmodulin, [U-100% 13C; U-100% 15N], 1.5 mM; IQ motif peptide of NaV1.2 1.5 mM; CALCIUM ION 3.3 mM; imidazole, [U-2H], 10 mM; potassium chloride 100 mM; sodium azide 0.01%; EDTA, [U-2H], 50 uM; D2O 100%
sample_conditions_1: pH: 6.8; pressure: 1 atm; temperature: 298 K
Experiments:
| Name | Sample | Sample state | Sample conditions |
|---|---|---|---|
| 2D 1H-15N HSQC | sample_1 | isotropic | sample_conditions_1 |
| 2D 1H-13C HMQC | sample_2 | isotropic | sample_conditions_1 |
| 3D HNCA | sample_1 | isotropic | sample_conditions_1 |
| 3D HN(CO)CA | sample_1 | isotropic | sample_conditions_1 |
| 3D HNCACB | sample_1 | isotropic | sample_conditions_1 |
| 3D HN(CO)CACB | sample_1 | isotropic | sample_conditions_1 |
| 3D HNCO | sample_1 | isotropic | sample_conditions_1 |
| 3D HN(CA)CO | sample_1 | isotropic | sample_conditions_1 |
| 3D C(CO)NH | sample_1 | isotropic | sample_conditions_1 |
| 3D H(CCO)NH | sample_1 | isotropic | sample_conditions_1 |
| 3D 1H-15N NOESY | sample_1 | isotropic | sample_conditions_1 |
| 3D 1H-13C NOESY | sample_2 | isotropic | sample_conditions_1 |
| 3D H(C)CH-TOCSY | sample_2 | isotropic | sample_conditions_1 |
| 2D 1H-15N HSQC | sample_1 | isotropic | sample_conditions_1 |
| 2D 1H-13C HMQC | sample_2 | isotropic | sample_conditions_1 |
| 3D 1H-13C edited, 12C filtered NOESY | sample_2 | isotropic | sample_conditions_1 |
| 2D 1H-1H doubly 12C,14N filtered NOESY | sample_1 | isotropic | sample_conditions_1 |
| 2D 1H-1H 12C,14N filtered TOCSY | sample_1 | isotropic | sample_conditions_1 |
| 2D 1H-1H 12C filtered NOESY | sample_2 | isotropic | sample_conditions_1 |
| 2D 1H-1H 12C,14N filtered TOCSY | sample_2 | isotropic | sample_conditions_1 |
Software:
TOPSPIN v2.1, Bruker Biospin - collection
SPARKY v3.115, Goddard - chemical shift assignment, data analysis, peak picking
NMRPipe, Delaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax - data analysis, processing
Analysis v2.1.5, CCPN - chemical shift assignment, data analysis, peak picking
ARIA v2.3, Linge, O'Donoghue and Nilges - data analysis, structure solution
CNSSOLVE v1.21, Brunger, Adams, Clore, Gros, Nilges and Read - data analysis, refinement, structure solution
X-PLOR NIH v2.23, Schwieters, Kuszewski, Tjandra and Clore - data analysis, refinement, structure solution
ProcheckNMR v3.5.4, Laskowski and MacArthur - data analysis
NMR spectrometers:
- Bruker Avance II 500 MHz
- Bruker Avance II 800 MHz
Download simulated HSQC data in one of the following formats:
CSV: Backbone
or all simulated shifts
SPARKY: Backbone
or all simulated shifts