BMRB Entry 26976
Chem Shift validation: AVS_full
BMRB Entry DOI: doi:10.13018/BMR26976
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Title: AILV Methyl Resonance Assignments for G protein alpha i3 subunit in the GoLoco14-bound state PubMed: 28223697
Deposition date: 2016-12-17 Original release date: 2017-02-21
Authors: Toyama, Yuki; Kano, Hanaho; Mase, Yoko; Yokogawa, Mariko; Osawa, Masanori; Shimada, Ichio
Citation: Toyama, Yuki; Kano, Hanaho; Mase, Yoko; Yokogawa, Mariko; Osawa, Masanori; Shimada, Ichio. "Dynamic regulation of GDP binding to G proteins revealed by magnetic field dependent NMR relaxation analyses" Nat. Commun. 8, .-. (2017).
Assembly members:
G_protein_alpha_subunit_i3, polymer, 359 residues, Formula weight is not available
entity_GDP, non-polymer, 443.201 Da.
GoLoco14_(Regulator_of_G-protein_Signaling_14), polymer, 41 residues, Formula weight is not available
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
Entity Sequences (FASTA):
G_protein_alpha_subunit_i3: GPDDHMGCTLSAEDKAAVER
SKMIDRNLREDGEKAAKEVK
LLLLGAGESGKSTIVKQMKI
IHEDGYSEDECKQYKVVVYS
NTIQSIIAIIRAMGRLKIDF
GEAARADDARQLFVLAGSAE
EGVMTPELAGVIKRLWRDGG
VQACFSRSREYQLNDSASYY
LNDLDRISQSNYIPTQQDVL
RTRVKTTGIVETHFTFKDLY
FKMFDVGGQRSERKKWIHCF
EGVTAIIFCVALSDYDLVLA
EDEEMNRMHESMKLFDSICN
NKWFTETSIILFLNKKDLFE
EKIKRSPLTICYPEYTGSNT
YEEAAAYIQCQFEDLNRRKD
TKEIYTHFTCATDTKNVQFV
FDAVTDVIIKNNLKECGLY
GoLoco14_(Regulator_of_G-protein_Signaling_14): GPLGSDIEGLVELLNRVQSS
GAHDQRGLLRKEDLVLPEFL
Q
- assigned_chemical_shifts
| Data type | Count |
| 13C chemical shifts | 93 |
| 1H chemical shifts | 279 |
Additional metadata:
Assembly:
| Entity Assembly ID | Entity Name | Entity ID |
|---|---|---|
| 1 | G alphai 3 | 1 |
| 2 | GDP | 2 |
| 3 | GoLoco14 | 3 |
Entities:
Entity 1, G alphai 3 359 residues - Formula weight is not available
| 1 | GLY | PRO | ASP | ASP | HIS | MET | GLY | CYS | THR | LEU | ||||
| 2 | SER | ALA | GLU | ASP | LYS | ALA | ALA | VAL | GLU | ARG | ||||
| 3 | SER | LYS | MET | ILE | ASP | ARG | ASN | LEU | ARG | GLU | ||||
| 4 | ASP | GLY | GLU | LYS | ALA | ALA | LYS | GLU | VAL | LYS | ||||
| 5 | LEU | LEU | LEU | LEU | GLY | ALA | GLY | GLU | SER | GLY | ||||
| 6 | LYS | SER | THR | ILE | VAL | LYS | GLN | MET | LYS | ILE | ||||
| 7 | ILE | HIS | GLU | ASP | GLY | TYR | SER | GLU | ASP | GLU | ||||
| 8 | CYS | LYS | GLN | TYR | LYS | VAL | VAL | VAL | TYR | SER | ||||
| 9 | ASN | THR | ILE | GLN | SER | ILE | ILE | ALA | ILE | ILE | ||||
| 10 | ARG | ALA | MET | GLY | ARG | LEU | LYS | ILE | ASP | PHE | ||||
| 11 | GLY | GLU | ALA | ALA | ARG | ALA | ASP | ASP | ALA | ARG | ||||
| 12 | GLN | LEU | PHE | VAL | LEU | ALA | GLY | SER | ALA | GLU | ||||
| 13 | GLU | GLY | VAL | MET | THR | PRO | GLU | LEU | ALA | GLY | ||||
| 14 | VAL | ILE | LYS | ARG | LEU | TRP | ARG | ASP | GLY | GLY | ||||
| 15 | VAL | GLN | ALA | CYS | PHE | SER | ARG | SER | ARG | GLU | ||||
| 16 | TYR | GLN | LEU | ASN | ASP | SER | ALA | SER | TYR | TYR | ||||
| 17 | LEU | ASN | ASP | LEU | ASP | ARG | ILE | SER | GLN | SER | ||||
| 18 | ASN | TYR | ILE | PRO | THR | GLN | GLN | ASP | VAL | LEU | ||||
| 19 | ARG | THR | ARG | VAL | LYS | THR | THR | GLY | ILE | VAL | ||||
| 20 | GLU | THR | HIS | PHE | THR | PHE | LYS | ASP | LEU | TYR | ||||
| 21 | PHE | LYS | MET | PHE | ASP | VAL | GLY | GLY | GLN | ARG | ||||
| 22 | SER | GLU | ARG | LYS | LYS | TRP | ILE | HIS | CYS | PHE | ||||
| 23 | GLU | GLY | VAL | THR | ALA | ILE | ILE | PHE | CYS | VAL | ||||
| 24 | ALA | LEU | SER | ASP | TYR | ASP | LEU | VAL | LEU | ALA | ||||
| 25 | GLU | ASP | GLU | GLU | MET | ASN | ARG | MET | HIS | GLU | ||||
| 26 | SER | MET | LYS | LEU | PHE | ASP | SER | ILE | CYS | ASN | ||||
| 27 | ASN | LYS | TRP | PHE | THR | GLU | THR | SER | ILE | ILE | ||||
| 28 | LEU | PHE | LEU | ASN | LYS | LYS | ASP | LEU | PHE | GLU | ||||
| 29 | GLU | LYS | ILE | LYS | ARG | SER | PRO | LEU | THR | ILE | ||||
| 30 | CYS | TYR | PRO | GLU | TYR | THR | GLY | SER | ASN | THR | ||||
| 31 | TYR | GLU | GLU | ALA | ALA | ALA | TYR | ILE | GLN | CYS | ||||
| 32 | GLN | PHE | GLU | ASP | LEU | ASN | ARG | ARG | LYS | ASP | ||||
| 33 | THR | LYS | GLU | ILE | TYR | THR | HIS | PHE | THR | CYS | ||||
| 34 | ALA | THR | ASP | THR | LYS | ASN | VAL | GLN | PHE | VAL | ||||
| 35 | PHE | ASP | ALA | VAL | THR | ASP | VAL | ILE | ILE | LYS | ||||
| 36 | ASN | ASN | LEU | LYS | GLU | CYS | GLY | LEU | TYR |
Entity 2, GDP - C10 H15 N5 O11 P2 - 443.201 Da.
| 1 | GDP |
Entity 3, GoLoco14 41 residues - Formula weight is not available
| 1 | GLY | PRO | LEU | GLY | SER | ASP | ILE | GLU | GLY | LEU | ||||
| 2 | VAL | GLU | LEU | LEU | ASN | ARG | VAL | GLN | SER | SER | ||||
| 3 | GLY | ALA | HIS | ASP | GLN | ARG | GLY | LEU | LEU | ARG | ||||
| 4 | LYS | GLU | ASP | LEU | VAL | LEU | PRO | GLU | PHE | LEU | ||||
| 5 | GLN |
Samples:
sample_1: G protein alpha subunit i3, [U-2H, AILVproS 1H,13C], 270 uM; HEPES 20 mM; DTT 5 mM; GUANOSINE-5'-DIPHOSPHATE 500 uM; GoLoco14 (Regulator of G-protein Signaling 14) 290 uM; DSS 100 uM; D2O 100%
sample_conditions_1: ionic strength: 20 mM; pH: 7; pressure: 1 atm; temperature: 293 K
Experiments:
| Name | Sample | Sample state | Sample conditions |
|---|---|---|---|
| 2D 1H-13C HMQC | sample_1 | isotropic | sample_conditions_1 |
| 3D 1H-13C NOESY aliphatic | sample_1 | isotropic | sample_conditions_1 |
Software:
TOPSPIN vv3.1, Bruker Biospin - collection, processing
SPARKY v3.114, Goddard - chemical shift assignment, data analysis, peak picking
NMR spectrometers:
- Bruker Avance 800 MHz