BMRB Entry 12003
Chem Shift validation: AVS_full
BMRB Entry DOI: doi:10.13018/BMR12003
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Title: ILMV methyl assignments for the neuronal SNARE complex PubMed: 27988858
Deposition date: 2016-09-12 Original release date: 2017-03-07
Authors: Pan, Yun-Zu; Quade, Bradley; Brewer, Kyle; Graham, Bim; Rizo, Josep; Swarbrick, James; Szabo, Monika
Citation: Pan, Yun-Zu; Quade, Bradley; Brewer, Kyle; Szabo, Monika; Swarbrick, James; Graham, Bim; Rizo, Josep. "Sequence-specific assignment of methyl groups from the neuronal SNARE complex using lanthanide-induced pseudocontact shifts" J. Biomol. NMR 66, 281-293 (2016).
Assembly members:
SNAP-25N, polymer, 75 residues, Formula weight is not available
SNAP-25C, polymer, 66 residues, Formula weight is not available
synaptobrevin_2, polymer, 67 residues, Formula weight is not available
syntaxin-1A, polymer, 65 residues, Formula weight is not available
Natural source: Common Name: Rat Taxonomy ID: 10116 Superkingdom: Eukaryota Kingdom: Metazoa Genus/species: Rattus norvegicus
Experimental source: Production method: recombinant technology Host organism: Escherichia coli Vector: pET28a
Entity Sequences (FASTA):
SNAP-25N: GSWLEEMQRRADQLADESLE
STRRMLQLVEESKDAGIRTL
VMLDEQGEQLDRVEEGMNHI
NQDMKEAEKNLKDLG
SNAP-25C: GSWARENEMDENLEQVSGII
GNLRHMALDMGNEIDTQNRQ
IDRIMEKADSNKTRIDEANQ
RATKML
synaptobrevin_2: GSNRRLQQTQAQVDEVVDIM
RVNVDKVLERDQKLSELDDR
ADALQAGASQFETSAAKLKR
KYWWKNL
syntaxin-1A: GSALSEIETRHSEIIKLENS
IRELHDMFMDMAMLVESQGE
MIDRIEYNVEHAVDYVERAV
SDTKK
| Data type | Count |
| 13C chemical shifts | 111 |
| 1H chemical shifts | 333 |
Additional metadata:
Assembly:
| Entity Assembly ID | Entity Name | Entity ID |
|---|---|---|
| 1 | SNAP-25 N-terminal SNARE motif | 1 |
| 2 | SNAP-25 C-terminal SNARE motif | 2 |
| 3 | synaptobrevin-2 SNARE motif | 3 |
| 4 | syntaxin-1A SNARE motif | 4 |
Entities:
Entity 1, SNAP-25 N-terminal SNARE motif 75 residues - Formula weight is not available
Residues 4-75 correspond to residues 11-82 of rat SNAP-25A. GSW at the N-terminus arise from the expression vector.
| 1 | GLY | SER | TRP | LEU | GLU | GLU | MET | GLN | ARG | ARG | ||||
| 2 | ALA | ASP | GLN | LEU | ALA | ASP | GLU | SER | LEU | GLU | ||||
| 3 | SER | THR | ARG | ARG | MET | LEU | GLN | LEU | VAL | GLU | ||||
| 4 | GLU | SER | LYS | ASP | ALA | GLY | ILE | ARG | THR | LEU | ||||
| 5 | VAL | MET | LEU | ASP | GLU | GLN | GLY | GLU | GLN | LEU | ||||
| 6 | ASP | ARG | VAL | GLU | GLU | GLY | MET | ASN | HIS | ILE | ||||
| 7 | ASN | GLN | ASP | MET | LYS | GLU | ALA | GLU | LYS | ASN | ||||
| 8 | LEU | LYS | ASP | LEU | GLY |
Entity 2, SNAP-25 C-terminal SNARE motif 66 residues - Formula weight is not available
Residues 4-66 correspond to residues 141-203 of rat SNAP-25A. GSW at the N-terminus arise from the expression vector.
| 1 | GLY | SER | TRP | ALA | ARG | GLU | ASN | GLU | MET | ASP | ||||
| 2 | GLU | ASN | LEU | GLU | GLN | VAL | SER | GLY | ILE | ILE | ||||
| 3 | GLY | ASN | LEU | ARG | HIS | MET | ALA | LEU | ASP | MET | ||||
| 4 | GLY | ASN | GLU | ILE | ASP | THR | GLN | ASN | ARG | GLN | ||||
| 5 | ILE | ASP | ARG | ILE | MET | GLU | LYS | ALA | ASP | SER | ||||
| 6 | ASN | LYS | THR | ARG | ILE | ASP | GLU | ALA | ASN | GLN | ||||
| 7 | ARG | ALA | THR | LYS | MET | LEU |
Entity 3, synaptobrevin-2 SNARE motif 67 residues - Formula weight is not available
Residues 3-67 correspond to residues 29-93 of rat synaptobrevin 2. GS at the N-terminus arises from the expression vector.
| 1 | GLY | SER | ASN | ARG | ARG | LEU | GLN | GLN | THR | GLN | ||||
| 2 | ALA | GLN | VAL | ASP | GLU | VAL | VAL | ASP | ILE | MET | ||||
| 3 | ARG | VAL | ASN | VAL | ASP | LYS | VAL | LEU | GLU | ARG | ||||
| 4 | ASP | GLN | LYS | LEU | SER | GLU | LEU | ASP | ASP | ARG | ||||
| 5 | ALA | ASP | ALA | LEU | GLN | ALA | GLY | ALA | SER | GLN | ||||
| 6 | PHE | GLU | THR | SER | ALA | ALA | LYS | LEU | LYS | ARG | ||||
| 7 | LYS | TYR | TRP | TRP | LYS | ASN | LEU |
Entity 4, syntaxin-1A SNARE motif 65 residues - Formula weight is not available
Residues 3-65 correspond to residues 191-253 of rat syntaxin-1. GS at the N-terminus arises from the expression vector.
| 1 | GLY | SER | ALA | LEU | SER | GLU | ILE | GLU | THR | ARG | ||||
| 2 | HIS | SER | GLU | ILE | ILE | LYS | LEU | GLU | ASN | SER | ||||
| 3 | ILE | ARG | GLU | LEU | HIS | ASP | MET | PHE | MET | ASP | ||||
| 4 | MET | ALA | MET | LEU | VAL | GLU | SER | GLN | GLY | GLU | ||||
| 5 | MET | ILE | ASP | ARG | ILE | GLU | TYR | ASN | VAL | GLU | ||||
| 6 | HIS | ALA | VAL | ASP | TYR | VAL | GLU | ARG | ALA | VAL | ||||
| 7 | SER | ASP | THR | LYS | LYS |
Samples:
sample_1: SNAP-25N, [ILMV-13CH3; U-2H], 30 uM; SNAP-25C 30 uM; synaptobrevin 2 30 uM; syntaxin-1A 30 uM; sodium chloride 200 mM; TRIS 20 mM; H2O 90%; D2O, [U-2H], 10%
sample_2: SNAP-25N 30 uM; SNAP-25C, [ILMV-13CH3; U-2H], 30 uM; synaptobrevin 2 30 uM; syntaxin-1A 30 uM; sodium chloride 200 mM; TRIS 20 mM; H2O 90%; D2O, [U-2H], 10%
sample_3: SNAP-25N 30 uM; SNAP-25C 30 uM; synaptobrevin 2, [ILMV-13CH3; U-2H], 30 uM; syntaxin-1A 30 uM; sodium chloride 200 mM; TRIS 20 mM; H2O 90%; D2O, [U-2H], 10%
sample_4: SNAP-25N 30 uM; SNAP-25C 30 uM; synaptobrevin 2 30 uM; syntaxin-1A, [ILMV-13CH3; U-2H], 30 uM; sodium chloride 200 mM; TRIS 20 mM; H2O 90%; D2O, [U-2H], 10%
sample_conditions_1: ionic strength: 220 mM; pH: 7.4; pressure: 1 atm; temperature: 298 K
Experiments:
| Name | Sample | Sample state | Sample conditions |
|---|---|---|---|
| 2D 1H-13C HMQC | sample_1 | isotropic | sample_conditions_1 |
| 2D 1H-13C HMQC | sample_2 | isotropic | sample_conditions_1 |
| 2D 1H-13C HMQC | sample_3 | isotropic | sample_conditions_1 |
| 2D 1H-13C HMQC | sample_4 | isotropic | sample_conditions_1 |
Software:
NMRPipe, Delaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax - processing
NMRView, Johnson, One Moon Scientific - data analysis
NUMBAT, Christophe Schmitz - PCS analysis
NMR spectrometers:
- Agilent DD2 800 MHz
- Agilent DD2 600 MHz