BMRB Entry 30543
Chem Shift validation: AVS_anomalous, AVS_full, LACS
BMRB Entry DOI: doi:10.13018/BMR30543
NMR-STAR file interactive viewer.
NMR-STAR v3 text file.
NMR-STAR v2.1 text file (deprecated)
XML gzip file.
RDF gzip file.
All files associated with the entry
Title: NMR solution structure of the homodimeric, autoinhibited state of the CARD9 CARD and first coiled-coil PubMed: 31296852
Deposition date: 2018-11-13 Original release date: 2019-07-12
Authors: Holliday, M.; Fairbrother, W.; Dueber, E.
Citation: Holliday, M.; Witt, A.; Rodriguez Gama, A.; Walters, B.; Arthur, C.; Halfmann, R.; Rohou, A.; Dueber, E.; Fairbrother, W.. "Structures of autoinhibited and polymerized forms of CARD9 reveal mechanisms of CARD9 and CARD11 activation" Nat. Commun. 10, 3070-3070 (2019).
Assembly members:
entity_1, polymer, 142 residues, 16116.458 Da.
entity_ZN, non-polymer, 65.409 Da.
Natural source: Common Name: Human Taxonomy ID: 9606 Superkingdom: Eukaryota Kingdom: Metazoa Genus/species: Homo sapiens
Experimental source: Production method: recombinant technology Host organism: Escherichia coli BL21(DE3)
Entity Sequences (FASTA):
entity_1: GSDYENDDECWSVLEGFRVT
LTSVIDPSRITPYLRQCKVL
NPDDEEQVLSDPNLVIRKRK
VGVLLDILQRTGHKGYVAFL
ESLELYYPQLYKKVTGKEPA
RVFSMIIDASGESGLTQLLM
TEVMKLQKKVQDLTALLSSK
DD
- assigned_chemical_shifts
- spectral_peak_list
| Data type | Count |
| 13C chemical shifts | 1038 |
| 15N chemical shifts | 270 |
| 1H chemical shifts | 1149 |
Additional metadata:
Assembly:
| Entity Assembly ID | Entity Name | Entity ID |
|---|---|---|
| 1 | entity_1, 1 | 1 |
| 2 | entity_1, 2 | 1 |
| 3 | entity_2, 1 | 2 |
| 4 | entity_2, 2 | 2 |
Entities:
Entity 1, entity_1, 1 142 residues - 16116.458 Da.
| 1 | GLY | SER | ASP | TYR | GLU | ASN | ASP | ASP | GLU | CYS | ||||
| 2 | TRP | SER | VAL | LEU | GLU | GLY | PHE | ARG | VAL | THR | ||||
| 3 | LEU | THR | SER | VAL | ILE | ASP | PRO | SER | ARG | ILE | ||||
| 4 | THR | PRO | TYR | LEU | ARG | GLN | CYS | LYS | VAL | LEU | ||||
| 5 | ASN | PRO | ASP | ASP | GLU | GLU | GLN | VAL | LEU | SER | ||||
| 6 | ASP | PRO | ASN | LEU | VAL | ILE | ARG | LYS | ARG | LYS | ||||
| 7 | VAL | GLY | VAL | LEU | LEU | ASP | ILE | LEU | GLN | ARG | ||||
| 8 | THR | GLY | HIS | LYS | GLY | TYR | VAL | ALA | PHE | LEU | ||||
| 9 | GLU | SER | LEU | GLU | LEU | TYR | TYR | PRO | GLN | LEU | ||||
| 10 | TYR | LYS | LYS | VAL | THR | GLY | LYS | GLU | PRO | ALA | ||||
| 11 | ARG | VAL | PHE | SER | MET | ILE | ILE | ASP | ALA | SER | ||||
| 12 | GLY | GLU | SER | GLY | LEU | THR | GLN | LEU | LEU | MET | ||||
| 13 | THR | GLU | VAL | MET | LYS | LEU | GLN | LYS | LYS | VAL | ||||
| 14 | GLN | ASP | LEU | THR | ALA | LEU | LEU | SER | SER | LYS | ||||
| 15 | ASP | ASP |
Entity 2, entity_2, 1 - Zn - 65.409 Da.
| 1 | ZN |
Samples:
sample_1: CARD9_2-142, [U-13C; U-15N], 1 mM; Zinc 1 mM; HEPES 50 mM; NaCl 300 mM; TCEP 0.5 mM
sample_2: CARD9_2-142, [U-13C; U-15N], 1 mM; Zinc 1 mM; HEPES 50 mM; NaCl 300 mM; TCEP 0.5 mM
sample_3: CARD9_2-142, [U-13C; U-15N; U-2H], 1 mM; Zinc 1 mM; HEPES 50 mM; NaCl 300 mM; TCEP 0.5 mM
sample_4: CARD9_2-142, [U-13C; U-15N], 1 mM; Zinc 2 mM; unlabeled CARD9_2-142 1 mM; HEPES 50 mM; NaCl 300 mM; TCEP 0.5 mM
sample_5: CARD9_2-142, [U-13C; U-15N], 1 mM; Zinc 1 mM; filamentous Pf1 bacteriophage 14 mg/mL; HEPES 50 mM; NaCl 900 mM; TCEP 0.5 mM
sample_conditions_1: ionic strength: 300 mM; pH: 7.0; pressure: 1 atm; temperature: 310 K
sample_conditions_2: ionic strength: 900 mM; pH: 7.0; pressure: 1 atm; temperature: 310 K
Experiments:
| Name | Sample | Sample state | Sample conditions |
|---|---|---|---|
| 2D 1H-15N TROSY | sample_3 | isotropic | sample_conditions_1 |
| 2D 1H-13C HSQC aliphatic | sample_2 | isotropic | sample_conditions_1 |
| 2D 1H-13C TROSY aromatic | sample_2 | isotropic | sample_conditions_1 |
| 3D HNCA | sample_3 | isotropic | sample_conditions_1 |
| 3D HNCB | sample_3 | isotropic | sample_conditions_1 |
| 3D (H)CC(CO)NH | sample_1 | isotropic | sample_conditions_1 |
| 3D 1H-15N NOESY | sample_1 | isotropic | sample_conditions_1 |
| 3D HN(CA)CO | sample_3 | isotropic | sample_conditions_1 |
| 3D HNCO | sample_3 | isotropic | sample_conditions_1 |
| 3D CBCA(CO)NH | sample_3 | isotropic | sample_conditions_1 |
| 3D HNCA | sample_1 | isotropic | sample_conditions_1 |
| 3D CBCA(CO)NH | sample_3 | isotropic | sample_conditions_1 |
| 3D HCCH-TOCSY | sample_2 | isotropic | sample_conditions_1 |
| 3D HCCH-COSY | sample_2 | isotropic | sample_conditions_1 |
| 3D 1H-13C NOESY aliphatic | sample_1 | isotropic | sample_conditions_1 |
| 3D 1H-13C NOESY aliphatic | sample_2 | isotropic | sample_conditions_1 |
| 3D 1H-13C NOESY aromatic | sample_2 | isotropic | sample_conditions_1 |
| 3D HNCO | sample_1 | isotropic | sample_conditions_1 |
| 3D 1H-13C interNOESY aliphatic | sample_4 | isotropic | sample_conditions_1 |
| 2D IPAP 1H-15N TROSY | sample_5 | anisotropic | sample_conditions_2 |
Software:
CNS v1.2, Brunger A. T. et.al. - refinement
CYANA v3.97, Guntert, Mumenthaler and Wuthrich - refinement, structure calculation
Analysis v2.42, CCPN - chemical shift assignment, peak picking
NMR spectrometers:
- Bruker Avance 800 MHz
- Bruker Avance 600 MHz
Download simulated HSQC data in one of the following formats:
CSV: Backbone
or all simulated shifts
SPARKY: Backbone
or all simulated shifts