BMRB Entry 51445
Chem Shift validation: AVS_full
BMRB Entry DOI: doi:10.13018/BMR51445
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Title: Chemical shift assignments for alanine and valine methyls of Robo1-Ig1-2-LnLoop PubMed: 36042257
Deposition date: 2022-05-16 Original release date: 2022-06-14
Authors: Williams, Robert; Huang, Chin; Moremen, Kelley; Amster, I.; Prestegard, James
Citation: Williams, Robert; Huang, Chin; Moremen, Kelley; Amster, I.; Prestegard, James. "NMR analysis suggests the terminal domains of Robo1 remain extended but are rigidified in the presence of heparan sulfate" Sci. Rep. 12, 14769-14769 (2022).
Assembly members:
entity_1, polymer, 222 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: mammalian HEK Vector: pGEn2
Entity Sequences (FASTA):
entity_1: GSRLRQEDFPPRIVEHPSDL
IVSKGEPATLNCKAEGRPTP
TIEWYKGGERVETDKDDPRS
HRMLLPSGSLFFLRIVHGRK
SRPDEGVYVCVARNYLGEAV
SHNASLEVAILRDDFRQNPS
DVMVAVGEPAVMECQPPRGH
PEPTISWKKDGSPLDDKDER
ITIRCADTNNDGAYEGDELC
KLMITYTRKSDAGKYVCVGT
NMVGERESEVAELTVLERPS
FV
- assigned_chemical_shifts
| Data type | Count |
| 13C chemical shifts | 29 |
| 1H chemical shifts | 87 |
Additional metadata:
Assembly:
| Entity Assembly ID | Entity Name | Entity ID |
|---|---|---|
| 1 | Robo1-Ig1-2-LnLoop | 1 |
Entities:
Entity 1, Robo1-Ig1-2-LnLoop 222 residues - Formula weight is not available
Numbering is consistent with uniprot Q9YN67 from 58-221. Then a 16 residue lanthanide binding tag replaces residues 222 and 223, resulting in the following sequence numbers differing from Q9YN67 by +14.
| 1 | GLY | SER | ARG | LEU | ARG | GLN | GLU | ASP | PHE | PRO | ||||
| 2 | PRO | ARG | ILE | VAL | GLU | HIS | PRO | SER | ASP | LEU | ||||
| 3 | ILE | VAL | SER | LYS | GLY | GLU | PRO | ALA | THR | LEU | ||||
| 4 | ASN | CYS | LYS | ALA | GLU | GLY | ARG | PRO | THR | PRO | ||||
| 5 | THR | ILE | GLU | TRP | TYR | LYS | GLY | GLY | GLU | ARG | ||||
| 6 | VAL | GLU | THR | ASP | LYS | ASP | ASP | PRO | ARG | SER | ||||
| 7 | HIS | ARG | MET | LEU | LEU | PRO | SER | GLY | SER | LEU | ||||
| 8 | PHE | PHE | LEU | ARG | ILE | VAL | HIS | GLY | ARG | LYS | ||||
| 9 | SER | ARG | PRO | ASP | GLU | GLY | VAL | TYR | VAL | CYS | ||||
| 10 | VAL | ALA | ARG | ASN | TYR | LEU | GLY | GLU | ALA | VAL | ||||
| 11 | SER | HIS | ASN | ALA | SER | LEU | GLU | VAL | ALA | ILE | ||||
| 12 | LEU | ARG | ASP | ASP | PHE | ARG | GLN | ASN | PRO | SER | ||||
| 13 | ASP | VAL | MET | VAL | ALA | VAL | GLY | GLU | PRO | ALA | ||||
| 14 | VAL | MET | GLU | CYS | GLN | PRO | PRO | ARG | GLY | HIS | ||||
| 15 | PRO | GLU | PRO | THR | ILE | SER | TRP | LYS | LYS | ASP | ||||
| 16 | GLY | SER | PRO | LEU | ASP | ASP | LYS | ASP | GLU | ARG | ||||
| 17 | ILE | THR | ILE | ARG | CYS | ALA | ASP | THR | ASN | ASN | ||||
| 18 | ASP | GLY | ALA | TYR | GLU | GLY | ASP | GLU | LEU | CYS | ||||
| 19 | LYS | LEU | MET | ILE | THR | TYR | THR | ARG | LYS | SER | ||||
| 20 | ASP | ALA | GLY | LYS | TYR | VAL | CYS | VAL | GLY | THR | ||||
| 21 | ASN | MET | VAL | GLY | GLU | ARG | GLU | SER | GLU | VAL | ||||
| 22 | ALA | GLU | LEU | THR | VAL | LEU | GLU | ARG | PRO | SER | ||||
| 23 | PHE | VAL |
Samples:
sample_1: Robo1-Ig1-2-LnLoop, [U-13C; 49% HB-Ala; 99% HG-Val], 300 uM; D2O, [U-99% 2H], 10%; TRIS 25 mM; NaCl 100 mM
sample_conditions_1: ionic strength: 100 mM; pH: 7.4; pressure: 1 atm; temperature: 298 K
Experiments:
| Name | Sample | Sample state | Sample conditions |
|---|---|---|---|
| 2D 13C-detected HSQC | sample_1 | isotropic | sample_conditions_1 |
| 3D 13C-detected NOESY | sample_1 | isotropic | sample_conditions_1 |
| 2D 13C-detected TOCSY | sample_1 | isotropic | sample_conditions_1 |
Software:
ASSIGN_SLPGUI v2.4.6 - chemical shift assignment
NMR spectrometers:
- Bruker AVANCE NEO 900 MHz