BMRB Entry 50530
Chem Shift validation: AVS_full
BMRB Entry DOI: doi:10.13018/BMR50530
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Title: Methyl assignments of recombinant wild-type human alpha1-antitrypsin PubMed: 33311470
Deposition date: 2020-10-28 Original release date: 2021-04-22
Authors: Jagger, Alistair; Waudby, Christopher; Irving, James; Christodoulou, John; Lomas, David
Citation: Jagger, Alistair; Waudby, Christopher; Irving, James; Christodoulou, John; Lomas, David. "High-resolution ex vivo NMR spectroscopy of human Z a1-antitrypsin" Nat. Commun. 11, 6371-6371 (2020).
Assembly members:
entity_1, polymer, 404 residues, Formula weight is not available
Natural source: Common Name: E. coli Taxonomy ID: 562 Superkingdom: Bacteria Kingdom: not available Genus/species: Escherichia coli
Experimental source: Production method: recombinant technology Host organism: Escherichia coli Vector: pQE-30
- assigned_chemical_shifts
| Data type | Count |
| 13C chemical shifts | 196 |
| 1H chemical shifts | 588 |
Additional metadata:
Assembly:
| Entity Assembly ID | Entity Name | Entity ID |
|---|---|---|
| 1 | AAT | 1 |
Entities:
Entity 1, AAT 404 residues - Formula weight is not available
Residues 1-11 represent a non-native affinity tag. Subsequent residue numbers are offset by 10 from that of the mature protein.
| 1 | MET | ARG | GLY | SER | HIS | HIS | HIS | HIS | HIS | HIS | ||||
| 2 | THR | ASP | PRO | GLN | GLY | ASP | ALA | ALA | GLN | LYS | ||||
| 3 | THR | ASP | THR | SER | HIS | HIS | ASP | GLN | ASP | HIS | ||||
| 4 | PRO | THR | PHE | ASN | LYS | ILE | THR | PRO | ASN | LEU | ||||
| 5 | ALA | GLU | PHE | ALA | PHE | SER | LEU | TYR | ARG | GLN | ||||
| 6 | LEU | ALA | HIS | GLN | SER | ASN | SER | THR | ASN | ILE | ||||
| 7 | PHE | PHE | SER | PRO | VAL | SER | ILE | ALA | THR | ALA | ||||
| 8 | PHE | ALA | MET | LEU | SER | LEU | GLY | THR | LYS | ALA | ||||
| 9 | ASP | THR | HIS | ASP | GLU | ILE | LEU | GLU | GLY | LEU | ||||
| 10 | ASN | PHE | ASN | LEU | THR | GLU | ILE | PRO | GLU | ALA | ||||
| 11 | GLN | ILE | HIS | GLU | GLY | PHE | GLN | GLU | LEU | LEU | ||||
| 12 | ARG | THR | LEU | ASN | GLN | PRO | ASP | SER | GLN | LEU | ||||
| 13 | GLN | LEU | THR | THR | GLY | ASN | GLY | LEU | PHE | LEU | ||||
| 14 | SER | GLU | GLY | LEU | LYS | LEU | VAL | ASP | LYS | PHE | ||||
| 15 | LEU | GLU | ASP | VAL | LYS | LYS | LEU | TYR | HIS | SER | ||||
| 16 | GLU | ALA | PHE | THR | VAL | ASN | PHE | GLY | ASP | THR | ||||
| 17 | GLU | GLU | ALA | LYS | LYS | GLN | ILE | ASN | ASP | TYR | ||||
| 18 | VAL | GLU | LYS | GLY | THR | GLN | GLY | LYS | ILE | VAL | ||||
| 19 | ASP | LEU | VAL | LYS | GLU | LEU | ASP | ARG | ASP | THR | ||||
| 20 | VAL | PHE | ALA | LEU | VAL | ASN | TYR | ILE | PHE | PHE | ||||
| 21 | LYS | GLY | LYS | TRP | GLU | ARG | PRO | PHE | GLU | VAL | ||||
| 22 | LYS | ASP | THR | GLU | GLU | GLU | ASP | PHE | HIS | VAL | ||||
| 23 | ASP | GLN | VAL | THR | THR | VAL | LYS | VAL | PRO | MET | ||||
| 24 | MET | LYS | ARG | LEU | GLY | MET | PHE | ASN | ILE | GLN | ||||
| 25 | HIS | CYS | LYS | LYS | LEU | SER | SER | TRP | VAL | LEU | ||||
| 26 | LEU | MET | LYS | TYR | LEU | GLY | ASN | ALA | THR | ALA | ||||
| 27 | ILE | PHE | PHE | LEU | PRO | ASP | GLU | GLY | LYS | LEU | ||||
| 28 | GLN | HIS | LEU | GLU | ASN | GLU | LEU | THR | HIS | ASP | ||||
| 29 | ILE | ILE | THR | LYS | PHE | LEU | GLU | ASN | GLU | ASP | ||||
| 30 | ARG | ARG | SER | ALA | SER | LEU | HIS | LEU | PRO | LYS | ||||
| 31 | LEU | SER | ILE | THR | GLY | THR | TYR | ASP | LEU | LYS | ||||
| 32 | SER | VAL | LEU | GLY | GLN | LEU | GLY | ILE | THR | LYS | ||||
| 33 | VAL | PHE | SER | ASN | GLY | ALA | ASP | LEU | SER | GLY | ||||
| 34 | VAL | THR | GLU | GLU | ALA | PRO | LEU | LYS | LEU | SER | ||||
| 35 | LYS | ALA | VAL | HIS | LYS | ALA | VAL | LEU | THR | ILE | ||||
| 36 | ASP | GLU | LYS | GLY | THR | GLU | ALA | ALA | GLY | ALA | ||||
| 37 | MET | PHE | LEU | GLU | ALA | ILE | PRO | MET | SER | ILE | ||||
| 38 | PRO | PRO | GLU | VAL | LYS | PHE | ASN | LYS | PRO | PHE | ||||
| 39 | VAL | PHE | LEU | MET | ILE | GLU | GLN | ASN | THR | LYS | ||||
| 40 | SER | PRO | LEU | PHE | MET | GLY | LYS | VAL | VAL | ASN | ||||
| 41 | PRO | THR | GLN | LYS |
Samples:
sample_1: alpha1-antitrypsin, [U-100% 2H; U-100% 15N; 13CH3-100% A(CB)I(CD1)L(CD2)M(CE)V(CG2)], 160 uM; sodium phosphate 25 mM; sodium chloride 50 mM; DTT 1 mM; EDTA 1 mM; sodium azide 0.02 % w/v
sample_2: alpha1-antitrypsin, [U-100% 2H; U-100% 15N; 13CH3-100% I(CD1)L(CD1/CD2)V(CG1/CG2)], 150 uM; sodium phosphate 25 mM; sodium chloride 50 mM; DTT 1 mM; EDTA 1 mM; sodium azide 0.02 % w/v
sample_3: alpha1-antitrypsin, [U-100% 2H; U-100% 15N; 13CH3-100% A(CB)I(CD1/CG2)L(CD1/CD2)V(CG1/CG2)], 165 uM; sodium phosphate 25 mM; sodium chloride 50 mM; DTT 1 mM; EDTA 1 mM; sodium azide 0.02 % w/v
sample_conditions_1: ionic strength: 400 mM; pH*: 7.6; pressure: 1 atm; temperature: 298 K
Experiments:
| Name | Sample | Sample state | Sample conditions |
|---|---|---|---|
| 2D 1H-13C HMQC | sample_1 | isotropic | sample_conditions_1 |
| 4D CC NOESY | sample_1 | isotropic | sample_conditions_1 |
| 2D 1H-13C HMQC | sample_2 | isotropic | sample_conditions_1 |
| 4D CC NOESY | sample_2 | isotropic | sample_conditions_1 |
| 2D 1H-13C HSQC | sample_3 | isotropic | sample_conditions_1 |
| 3D HMCMCG | sample_3 | isotropic | sample_conditions_1 |
| 3D HMCM(CG)CB | sample_3 | isotropic | sample_conditions_1 |
| 3D HMCM(CGCB)CA | sample_3 | isotropic | sample_conditions_1 |
| 3D HMCM(CG)CO | sample_3 | isotropic | sample_conditions_1 |
| 3D HMCM(CGCB)CO | sample_3 | isotropic | sample_conditions_1 |
| 3D HMCM(CGCBCA)CO | sample_3 | isotropic | sample_conditions_1 |
Software:
NMRPipe - processing
TOPSPIN - collection
CcpNMR - chemical shift assignment, peak picking
MATLAB - peak picking
SMILE - processing
NMR spectrometers:
- Bruker AVANCE III 900 MHz
- Bruker AVANCE III 600 MHz