BMRB Entry 19050

Title:
Structure of the C-domain of Calcium-saturated Calmodulin bound to the IQ motif of NaV1.2
Deposition date:
2013-02-21
Original release date:
2014-07-21
Authors:
Fowler, C.; Feldkamp, Michael; Yu, Liping; Shea, Madeline
Citation:

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).
PubMed: 28343066

Assembly members:

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:

Natural source:   Common Name: Ciliates   Taxonomy ID: 5888   Superkingdom: Eukaryota   Kingdom: not available   Genus/species: Paramecium tetraurelia

Experimental source:

Experimental source:   Production method: recombinant technology   Host organism: Escherichia coli   Vector: PT7-7

Entity Sequences (FASTA):

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

Data sets:
Data typeCount
13C chemical shifts314
15N chemical shifts78
1H chemical shifts606

Additional metadata:

  • Assembly
  • Samples and Experiments
  • Software
  • Spectrometers
  • Hide all

Assembly:

Entity Assembly IDEntity NameEntity ID
1C-domain of Calmodulin1
2IQ motif peptide of NaV1.22
3CALCIUM ION_13
4CALCIUM ION_23

Entities:

Entity 1, C-domain of Calmodulin 73 residues - 8416.369 Da.

1   METLYSGLUGLNASPSERGLUGLUGLULEU
2   ILEGLUALAPHELYSVALPHEASPARGASP
3   GLYASNGLYLEUILESERALAALAGLULEU
4   ARGHISVALMETTHRASNLEUGLYGLULYS
5   LEUTHRASPASPGLUVALASPGLUMETILE
6   ARGGLUALAASPILEASPGLYASPGLYHIS
7   ILEASNTYRGLUGLUPHEVALARGMETMET
8   VALSERLYS

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   LYSARGLYSGLNGLUGLUVALSERALAILE
2   VALILEGLNARGALATYRARGARGTYRLEU
3   LEULYSGLNLYSVALLYSLYS

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:

NameSampleSample stateSample conditions
2D 1H-15N HSQCsample_1isotropicsample_conditions_1
2D 1H-13C HMQCsample_2isotropicsample_conditions_1
3D HNCAsample_1isotropicsample_conditions_1
3D HN(CO)CAsample_1isotropicsample_conditions_1
3D HNCACBsample_1isotropicsample_conditions_1
3D HN(CO)CACBsample_1isotropicsample_conditions_1
3D HNCOsample_1isotropicsample_conditions_1
3D HN(CA)COsample_1isotropicsample_conditions_1
3D C(CO)NHsample_1isotropicsample_conditions_1
3D H(CCO)NHsample_1isotropicsample_conditions_1
3D 1H-15N NOESYsample_1isotropicsample_conditions_1
3D 1H-13C NOESYsample_2isotropicsample_conditions_1
3D H(C)CH-TOCSYsample_2isotropicsample_conditions_1
2D 1H-15N HSQCsample_1isotropicsample_conditions_1
2D 1H-13C HMQCsample_2isotropicsample_conditions_1
3D 1H-13C edited, 12C filtered NOESYsample_2isotropicsample_conditions_1
2D 1H-1H doubly 12C,14N filtered NOESYsample_1isotropicsample_conditions_1
2D 1H-1H 12C,14N filtered TOCSYsample_1isotropicsample_conditions_1
2D 1H-1H 12C filtered NOESYsample_2isotropicsample_conditions_1
2D 1H-1H 12C,14N filtered TOCSYsample_2isotropicsample_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

Related Database Links:

NCBI XP_001448363.1 NP_036779.1
UniProt P04775.1
AlphaFold P04775

Download HSQC peak lists in one of the following formats:
CSV: Backbone or all simulated peaks
SPARKY: Backbone or all simulated peaks