BMRB Entry 26584
Chem Shift validation: AVS_full, LACS
BMRB Entry DOI: doi:10.13018/BMR26584
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NMR-STAR v3 text file.
XML gzip file.
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Citation: Oyen, David; Fenwick, R Bryn; Aoto, Phillip; Stanfield, Robyn; Wilson, Ian; Dyson, H Jane; Wright, Peter. "Defining the Structural Basis for Allosteric Product Release from E. coli Dihydrofolate Reductase Using NMR Relaxation Dispersion" J. Am. Chem. Soc. 139, 11233-11240 (2017).
PubMed: 28737940
Assembly members:
dihydrofolate_reductase, polymer, 159 residues, 18028.2808 Da.
NADP NICOTINAMIDE-ADENINE-DINUCLEOTIDE PHOSPHATE, non-polymer, 743.405 Da.
FOLIC ACID, non-polymer, 441.397 Da.
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: pET22b
Entity Sequences (FASTA):
dihydrofolate_reductase: MISLIAALAVDRVIGMENAM
PWNLPADFAWFKRNTLNKPV
IMGRHTWESIGRPLPGRKNI
ILSSQPGTDDRVTWVKSVDE
AIAACGDVPEIMVIGGGRVY
EQFLPKAQKLYLTHIDAEVE
GDTHFPDYEPDDWESVFSEF
HDADAQNSHSYCFEILERR
- assigned_chemical_shifts
| Data type | Count |
| 13C chemical shifts | 229 |
| 15N chemical shifts | 139 |
| 1H chemical shifts | 139 |
Additional metadata:
Assembly:
| Entity Assembly ID | Entity Name | Entity ID |
|---|---|---|
| 1 | dihydrofolate reductase mutation (L28F) | 1 |
| 2 | NADP NICOTINAMIDE-ADENINE-DINUCLEOTIDE PHOSPHATE | 2 |
| 3 | FOLIC ACID | 3 |
Entities:
Entity 1, dihydrofolate reductase mutation (L28F) 159 residues - 18028.2808 Da.
| 1 | MET | ILE | SER | LEU | ILE | ALA | ALA | LEU | ALA | VAL | ||||
| 2 | ASP | ARG | VAL | ILE | GLY | MET | GLU | ASN | ALA | MET | ||||
| 3 | PRO | TRP | ASN | LEU | PRO | ALA | ASP | PHE | ALA | TRP | ||||
| 4 | PHE | LYS | ARG | ASN | THR | LEU | ASN | LYS | PRO | VAL | ||||
| 5 | ILE | MET | GLY | ARG | HIS | THR | TRP | GLU | SER | ILE | ||||
| 6 | GLY | ARG | PRO | LEU | PRO | GLY | ARG | LYS | ASN | ILE | ||||
| 7 | ILE | LEU | SER | SER | GLN | PRO | GLY | THR | ASP | ASP | ||||
| 8 | ARG | VAL | THR | TRP | VAL | LYS | SER | VAL | ASP | GLU | ||||
| 9 | ALA | ILE | ALA | ALA | CYS | GLY | ASP | VAL | PRO | GLU | ||||
| 10 | ILE | MET | VAL | ILE | GLY | GLY | GLY | ARG | VAL | TYR | ||||
| 11 | GLU | GLN | PHE | LEU | PRO | LYS | ALA | GLN | LYS | LEU | ||||
| 12 | TYR | LEU | THR | HIS | ILE | ASP | ALA | GLU | VAL | GLU | ||||
| 13 | GLY | ASP | THR | HIS | PHE | PRO | ASP | TYR | GLU | PRO | ||||
| 14 | ASP | ASP | TRP | GLU | SER | VAL | PHE | SER | GLU | PHE | ||||
| 15 | HIS | ASP | ALA | ASP | ALA | GLN | ASN | SER | HIS | SER | ||||
| 16 | TYR | CYS | PHE | GLU | ILE | LEU | GLU | ARG | ARG |
Entity 2, NADP NICOTINAMIDE-ADENINE-DINUCLEOTIDE PHOSPHATE - C21 H28 N7 O17 P3 - 743.405 Da.
| 1 | NAP |
Entity 3, FOLIC ACID - C19 H19 N7 O6 - 441.397 Da.
| 1 | FOL |
Samples:
sample_1: L28F ecDHFR, [U-100% 13C; U-100% 15N], 1 mM; Folate 18 mM; NADP+ 10 mM; ascorbic acid 5 mM; dithiothreitol 1 mM; KCl 25 mM; KPi 70 mM
sample_conditions_1: pH: 7.6; pressure: 1 atm; temperature: 300 K
Experiments:
| Name | Sample | Sample state | Sample conditions |
|---|---|---|---|
| 2D 1H-15N HSQC | sample_1 | isotropic | sample_conditions_1 |
| 3D CBCA(CO)NH | sample_1 | isotropic | sample_conditions_1 |
| 3D HNCACB | sample_1 | isotropic | sample_conditions_1 |
Software:
CcpNmr_Analysis v2.2, CCPN, Delaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax - chemical shift assignment, peak picking, processing
NMR spectrometers:
- Bruker Avance 800 MHz
- Bruker Avance 750 MHz
Download HSQC peak lists in one of the following formats:
CSV: Backbone
or all simulated peaks
SPARKY: Backbone
or all simulated peaks