Solution NMR Structure of the Ras-binding domain of Serine/threonine-protein kinase B-raf from Homo sapiens, Northeast Structural Genomics Consortium Target HR4694F
MGHHHHHHSH MPKSPQKPIV RVFLPNKQRT VVPARCGVTV RDSLKKALMM RGLIPECCAV YRIQDGEKKP IGWDTDISWL TGEELHVEVL ENVPL
Polymer type: polypeptide(L)
Total | 1H | 13C | 15N | |
---|---|---|---|---|
All | 88.8 % (1003 of 1130) | 89.0 % (528 of 593) | 88.3 % (391 of 443) | 89.4 % (84 of 94) |
Backbone | 88.3 % (489 of 554) | 88.3 % (166 of 188) | 88.2 % (246 of 279) | 88.5 % (77 of 87) |
Sidechain | 89.2 % (593 of 665) | 89.4 % (362 of 405) | 88.5 % (224 of 253) | 100.0 % (7 of 7) |
Aromatic | 62.2 % (46 of 74) | 62.2 % (23 of 37) | 60.0 % (21 of 35) | 100.0 % (2 of 2) |
Methyl | 98.1 % (104 of 106) | 96.2 % (51 of 53) | 100.0 % (53 of 53) |
1. HR4694F
MGHHHHHHSH MPKSPQKPIV RVFLPNKQRT VVPARCGVTV RDSLKKALMM RGLIPECCAV YRIQDGEKKP IGWDTDISWL TGEELHVEVL ENVPLSolvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 298 K, pH 4.5
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
1 | HR4694F | [U-100% 13C; U-100% 15N] | 0.84 mM | |
2 | ammonium acetate | natural abundance | 20 mM | |
3 | sodium chloride | natural abundance | 100 mM | |
4 | calcium chloride | natural abundance | 5 mM | |
5 | DTT | natural abundance | 10 mM | |
6 | sodium azide | natural abundance | 0.02 % | |
7 | DSS | natural abundance | 50 uM | |
8 | H2O | natural abundance | 90 % | |
9 | D2O | natural abundance | 10 % |
Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 298 K, pH 4.5
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
10 | HR4694F | [U-5% 13C; U-100% 15N] | 0.69 mM | |
11 | ammonium acetate | natural abundance | 20 mM | |
12 | sodium chloride | natural abundance | 100 mM | |
13 | calcium chloride | natural abundance | 5 mM | |
14 | DTT | natural abundance | 10 mM | |
15 | sodium azide | natural abundance | 0.02 % | |
16 | DSS | natural abundance | 50 uM | |
17 | H2O | natural abundance | 90 % | |
18 | D2O | natural abundance | 10 % |
Atom type | Mol. common name | Atom group | Value | Ref. method | Ref. type | Shift ratio |
---|---|---|---|---|---|---|
13C | DSS | methyl protons | 0.0 ppm | na | indirect | 0.2514495 |
1H | DSS | methyl protons | 0.0 ppm | internal | direct | 1.0 |
15N | DSS | methyl protons | 0.0 ppm | na | indirect | 0.1013291 |
Atom type | Mol. common name | Atom group | Value | Ref. method | Ref. type | Shift ratio |
---|---|---|---|---|---|---|
13C | DSS | methyl protons | 0.0 ppm | na | indirect | 0.2514495 |
1H | DSS | methyl protons | 0.0 ppm | internal | direct | 1.0 |
15N | DSS | methyl protons | 0.0 ppm | na | indirect | 0.1013291 |
Atom type | Mol. common name | Atom group | Value | Ref. method | Ref. type | Shift ratio |
---|---|---|---|---|---|---|
13C | DSS | methyl protons | 0.0 ppm | na | indirect | 0.2514495 |
1H | DSS | methyl protons | 0.0 ppm | internal | direct | 1.0 |
15N | DSS | methyl protons | 0.0 ppm | na | indirect | 0.1013291 |
Atom type | Mol. common name | Atom group | Value | Ref. method | Ref. type | Shift ratio |
---|---|---|---|---|---|---|
13C | DSS | methyl protons | 0.0 ppm | na | indirect | 0.2514495 |
1H | DSS | methyl protons | 0.0 ppm | internal | direct | 1.0 |
15N | DSS | methyl protons | 0.0 ppm | na | indirect | 0.1013291 |
Bruker Avance - 800 MHz 5-mm TXI cryoprobe
State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 298 K, pH 4.5
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
1 | HR4694F | [U-100% 13C; U-100% 15N] | 0.84 mM | |
2 | ammonium acetate | natural abundance | 20 mM | |
3 | sodium chloride | natural abundance | 100 mM | |
4 | calcium chloride | natural abundance | 5 mM | |
5 | DTT | natural abundance | 10 mM | |
6 | sodium azide | natural abundance | 0.02 % | |
7 | DSS | natural abundance | 50 uM | |
8 | H2O | natural abundance | 90 % | |
9 | D2O | natural abundance | 10 % |
Bruker Avance - 800 MHz 5-mm TXI cryoprobe
State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 298 K, pH 4.5
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
1 | HR4694F | [U-100% 13C; U-100% 15N] | 0.84 mM | |
2 | ammonium acetate | natural abundance | 20 mM | |
3 | sodium chloride | natural abundance | 100 mM | |
4 | calcium chloride | natural abundance | 5 mM | |
5 | DTT | natural abundance | 10 mM | |
6 | sodium azide | natural abundance | 0.02 % | |
7 | DSS | natural abundance | 50 uM | |
8 | H2O | natural abundance | 90 % | |
9 | D2O | natural abundance | 10 % |
Bruker Avance - 800 MHz 5-mm TXI cryoprobe
State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 298 K, pH 4.5
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
1 | HR4694F | [U-100% 13C; U-100% 15N] | 0.84 mM | |
2 | ammonium acetate | natural abundance | 20 mM | |
3 | sodium chloride | natural abundance | 100 mM | |
4 | calcium chloride | natural abundance | 5 mM | |
5 | DTT | natural abundance | 10 mM | |
6 | sodium azide | natural abundance | 0.02 % | |
7 | DSS | natural abundance | 50 uM | |
8 | H2O | natural abundance | 90 % | |
9 | D2O | natural abundance | 10 % |
Bruker Avance - 800 MHz 5-mm TXI cryoprobe
State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 298 K, pH 4.5
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
1 | HR4694F | [U-100% 13C; U-100% 15N] | 0.84 mM | |
2 | ammonium acetate | natural abundance | 20 mM | |
3 | sodium chloride | natural abundance | 100 mM | |
4 | calcium chloride | natural abundance | 5 mM | |
5 | DTT | natural abundance | 10 mM | |
6 | sodium azide | natural abundance | 0.02 % | |
7 | DSS | natural abundance | 50 uM | |
8 | H2O | natural abundance | 90 % | |
9 | D2O | natural abundance | 10 % |
Bruker Avance - 800 MHz 5-mm TXI cryoprobe
State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 298 K, pH 4.5
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
1 | HR4694F | [U-100% 13C; U-100% 15N] | 0.84 mM | |
2 | ammonium acetate | natural abundance | 20 mM | |
3 | sodium chloride | natural abundance | 100 mM | |
4 | calcium chloride | natural abundance | 5 mM | |
5 | DTT | natural abundance | 10 mM | |
6 | sodium azide | natural abundance | 0.02 % | |
7 | DSS | natural abundance | 50 uM | |
8 | H2O | natural abundance | 90 % | |
9 | D2O | natural abundance | 10 % |
Bruker Avance - 800 MHz 5-mm TXI cryoprobe
State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 298 K, pH 4.5
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
10 | HR4694F | [U-5% 13C; U-100% 15N] | 0.69 mM | |
11 | ammonium acetate | natural abundance | 20 mM | |
12 | sodium chloride | natural abundance | 100 mM | |
13 | calcium chloride | natural abundance | 5 mM | |
14 | DTT | natural abundance | 10 mM | |
15 | sodium azide | natural abundance | 0.02 % | |
16 | DSS | natural abundance | 50 uM | |
17 | H2O | natural abundance | 90 % | |
18 | D2O | natural abundance | 10 % |
Bruker Avance - 600 MHz 1.7-mm microcryoprobe
State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 298 K, pH 4.5
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
1 | HR4694F | [U-100% 13C; U-100% 15N] | 0.84 mM | |
2 | ammonium acetate | natural abundance | 20 mM | |
3 | sodium chloride | natural abundance | 100 mM | |
4 | calcium chloride | natural abundance | 5 mM | |
5 | DTT | natural abundance | 10 mM | |
6 | sodium azide | natural abundance | 0.02 % | |
7 | DSS | natural abundance | 50 uM | |
8 | H2O | natural abundance | 90 % | |
9 | D2O | natural abundance | 10 % |
Bruker Avance - 600 MHz 1.7-mm microcryoprobe
State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 298 K, pH 4.5
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
1 | HR4694F | [U-100% 13C; U-100% 15N] | 0.84 mM | |
2 | ammonium acetate | natural abundance | 20 mM | |
3 | sodium chloride | natural abundance | 100 mM | |
4 | calcium chloride | natural abundance | 5 mM | |
5 | DTT | natural abundance | 10 mM | |
6 | sodium azide | natural abundance | 0.02 % | |
7 | DSS | natural abundance | 50 uM | |
8 | H2O | natural abundance | 90 % | |
9 | D2O | natural abundance | 10 % |
Bruker Avance - 600 MHz 1.7-mm microcryoprobe
State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 298 K, pH 4.5
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
1 | HR4694F | [U-100% 13C; U-100% 15N] | 0.84 mM | |
2 | ammonium acetate | natural abundance | 20 mM | |
3 | sodium chloride | natural abundance | 100 mM | |
4 | calcium chloride | natural abundance | 5 mM | |
5 | DTT | natural abundance | 10 mM | |
6 | sodium azide | natural abundance | 0.02 % | |
7 | DSS | natural abundance | 50 uM | |
8 | H2O | natural abundance | 90 % | |
9 | D2O | natural abundance | 10 % |
Bruker Avance - 600 MHz 1.7-mm microcryoprobe
State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 298 K, pH 4.5
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
1 | HR4694F | [U-100% 13C; U-100% 15N] | 0.84 mM | |
2 | ammonium acetate | natural abundance | 20 mM | |
3 | sodium chloride | natural abundance | 100 mM | |
4 | calcium chloride | natural abundance | 5 mM | |
5 | DTT | natural abundance | 10 mM | |
6 | sodium azide | natural abundance | 0.02 % | |
7 | DSS | natural abundance | 50 uM | |
8 | H2O | natural abundance | 90 % | |
9 | D2O | natural abundance | 10 % |
Bruker Avance - 600 MHz 1.7-mm microcryoprobe
State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 298 K, pH 4.5
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
1 | HR4694F | [U-100% 13C; U-100% 15N] | 0.84 mM | |
2 | ammonium acetate | natural abundance | 20 mM | |
3 | sodium chloride | natural abundance | 100 mM | |
4 | calcium chloride | natural abundance | 5 mM | |
5 | DTT | natural abundance | 10 mM | |
6 | sodium azide | natural abundance | 0.02 % | |
7 | DSS | natural abundance | 50 uM | |
8 | H2O | natural abundance | 90 % | |
9 | D2O | natural abundance | 10 % |
Bruker Avance - 600 MHz 1.7-mm microcryoprobe
State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 298 K, pH 4.5
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
1 | HR4694F | [U-100% 13C; U-100% 15N] | 0.84 mM | |
2 | ammonium acetate | natural abundance | 20 mM | |
3 | sodium chloride | natural abundance | 100 mM | |
4 | calcium chloride | natural abundance | 5 mM | |
5 | DTT | natural abundance | 10 mM | |
6 | sodium azide | natural abundance | 0.02 % | |
7 | DSS | natural abundance | 50 uM | |
8 | H2O | natural abundance | 90 % | |
9 | D2O | natural abundance | 10 % |
Bruker Avance - 600 MHz 1.7-mm microcryoprobe
State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 298 K, pH 4.5
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
1 | HR4694F | [U-100% 13C; U-100% 15N] | 0.84 mM | |
2 | ammonium acetate | natural abundance | 20 mM | |
3 | sodium chloride | natural abundance | 100 mM | |
4 | calcium chloride | natural abundance | 5 mM | |
5 | DTT | natural abundance | 10 mM | |
6 | sodium azide | natural abundance | 0.02 % | |
7 | DSS | natural abundance | 50 uM | |
8 | H2O | natural abundance | 90 % | |
9 | D2O | natural abundance | 10 % |
Bruker Avance - 600 MHz 1.7-mm microcryoprobe
State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 298 K, pH 4.5
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
1 | HR4694F | [U-100% 13C; U-100% 15N] | 0.84 mM | |
2 | ammonium acetate | natural abundance | 20 mM | |
3 | sodium chloride | natural abundance | 100 mM | |
4 | calcium chloride | natural abundance | 5 mM | |
5 | DTT | natural abundance | 10 mM | |
6 | sodium azide | natural abundance | 0.02 % | |
7 | DSS | natural abundance | 50 uM | |
8 | H2O | natural abundance | 90 % | |
9 | D2O | natural abundance | 10 % |
Bruker Avance - 600 MHz 1.7-mm microcryoprobe
State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 298 K, pH 4.5
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
1 | HR4694F | [U-100% 13C; U-100% 15N] | 0.84 mM | |
2 | ammonium acetate | natural abundance | 20 mM | |
3 | sodium chloride | natural abundance | 100 mM | |
4 | calcium chloride | natural abundance | 5 mM | |
5 | DTT | natural abundance | 10 mM | |
6 | sodium azide | natural abundance | 0.02 % | |
7 | DSS | natural abundance | 50 uM | |
8 | H2O | natural abundance | 90 % | |
9 | D2O | natural abundance | 10 % |
Bruker Avance - 600 MHz 1.7-mm microcryoprobe
State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 298 K, pH 4.5
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
1 | HR4694F | [U-100% 13C; U-100% 15N] | 0.84 mM | |
2 | ammonium acetate | natural abundance | 20 mM | |
3 | sodium chloride | natural abundance | 100 mM | |
4 | calcium chloride | natural abundance | 5 mM | |
5 | DTT | natural abundance | 10 mM | |
6 | sodium azide | natural abundance | 0.02 % | |
7 | DSS | natural abundance | 50 uM | |
8 | H2O | natural abundance | 90 % | |
9 | D2O | natural abundance | 10 % |
Bruker Avance - 600 MHz 1.7-mm microcryoprobe
State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 298 K, pH 4.5
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
1 | HR4694F | [U-100% 13C; U-100% 15N] | 0.84 mM | |
2 | ammonium acetate | natural abundance | 20 mM | |
3 | sodium chloride | natural abundance | 100 mM | |
4 | calcium chloride | natural abundance | 5 mM | |
5 | DTT | natural abundance | 10 mM | |
6 | sodium azide | natural abundance | 0.02 % | |
7 | DSS | natural abundance | 50 uM | |
8 | H2O | natural abundance | 90 % | |
9 | D2O | natural abundance | 10 % |
Bruker Avance - 600 MHz 1.7-mm microcryoprobe
State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 298 K, pH 4.5
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
1 | HR4694F | [U-100% 13C; U-100% 15N] | 0.84 mM | |
2 | ammonium acetate | natural abundance | 20 mM | |
3 | sodium chloride | natural abundance | 100 mM | |
4 | calcium chloride | natural abundance | 5 mM | |
5 | DTT | natural abundance | 10 mM | |
6 | sodium azide | natural abundance | 0.02 % | |
7 | DSS | natural abundance | 50 uM | |
8 | H2O | natural abundance | 90 % | |
9 | D2O | natural abundance | 10 % |
Bruker Avance - 600 MHz 1.7-mm microcryoprobe
State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 298 K, pH 4.5
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
1 | HR4694F | [U-100% 13C; U-100% 15N] | 0.84 mM | |
2 | ammonium acetate | natural abundance | 20 mM | |
3 | sodium chloride | natural abundance | 100 mM | |
4 | calcium chloride | natural abundance | 5 mM | |
5 | DTT | natural abundance | 10 mM | |
6 | sodium azide | natural abundance | 0.02 % | |
7 | DSS | natural abundance | 50 uM | |
8 | H2O | natural abundance | 90 % | |
9 | D2O | natural abundance | 10 % |
Properties
Input source #1: NMR data (NEF) - Assigned chemical shifts, Distance restraints, Dihedral angle restraints | combined_17030_2l05.nef |
Input source #2: Coordindates | 2l05.cif |
Diamagnetism of the molecular assembly | True (excluding Oxygen atoms) |
Whether the assembly has a disulfide bond | None |
Whether the assembly has a other bond | None |
Whether the assembly contains a cyclic polymer | None |
Overall data processing status | Warning |
Disulfide bonds
NoneOther bonds (neither disulfide, covalent nor hydrogen bonds, e.g. Zinc–sulphur bond)
NoneNon-standard residues
NoneSequence alignments
--140-----150-------160-------170-------180-------190-------200-------210-------220-------230-- MGHHHHHHSHMPKSPQKPIVRVFLPNKQRTVVPARCGVTVRDSLKKALMMRGLIPECCAVYRIQDGEKKPIGWDTDISWLTGEELHVEVLENVPL ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||| MGHHHHHHSHMPKSPQKPIVRVFLPNKQRTVVPARCGVTVRDSLKKALMMRGLIPECCAVYRIQDGEKKPIGWDTDISWLTGEELHVEVLENVPL --------10--------20--------30--------40--------50--------60--------70--------80--------90-----
Chain assignments
Entity_assembly_ID (NMR) | Auth_asym_ID (model) | Length | Unmapped | Conflict | Sequence coverage (%) |
---|---|---|---|---|---|
A | A | 95 | 0 | 0 | 100.0 |
Content subtype: combined_17030_2l05.nef
Assigned chemical shifts
--140-----150-------160-------170-------180-------190-------200-------210-------220-------230-- MGHHHHHHSHMPKSPQKPIVRVFLPNKQRTVVPARCGVTVRDSLKKALMMRGLIPECCAVYRIQDGEKKPIGWDTDISWLTGEELHVEVLENVPL ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||| ..........MPKSPQKPIVRVFLPNKQRTVVPARCGVTVRDSLKKALMMRGLIPECCAVYRIQDGEKKPIGWDTDISWLTGEELHVEVLENVPL
Comp_index_ID | Comp_ID | Atom_ID | CS value (ppm) |
---|---|---|---|
153 | GLN | CD | 180.579 |
163 | ASN | CG | 178.447 |
165 | GLN | CD | 180.539 |
201 | GLN | CD | 179.709 |
229 | ASN | CG | 177.252 |
Atom group | # of target shifts | # of assigned shifts | Completeness (%) |
---|---|---|---|
1H chemical shifts | 593 | 534 | 90.1 |
13C chemical shifts | 443 | 389 | 87.8 |
15N chemical shifts | 100 | 89 | 89.0 |
Atom group | # of target shifts | # of assigned shifts | Completeness (%) |
---|---|---|---|
1H chemical shifts | 188 | 167 | 88.8 |
13C chemical shifts | 190 | 164 | 86.3 |
15N chemical shifts | 87 | 76 | 87.4 |
Atom group | # of target shifts | # of assigned shifts | Completeness (%) |
---|---|---|---|
1H chemical shifts | 405 | 367 | 90.6 |
13C chemical shifts | 253 | 225 | 88.9 |
15N chemical shifts | 13 | 13 | 100.0 |
Atom group | # of target shifts | # of assigned shifts | Completeness (%) |
---|---|---|---|
1H chemical shifts | 57 | 53 | 93.0 |
13C chemical shifts | 57 | 53 | 93.0 |
Atom group | # of target shifts | # of assigned shifts | Completeness (%) |
---|---|---|---|
1H chemical shifts | 37 | 23 | 62.2 |
13C chemical shifts | 35 | 21 | 60.0 |
15N chemical shifts | 2 | 2 | 100.0 |
Distance restraints
--140-----150-------160-------170-------180-------190-------200-------210-------220-------230-- MGHHHHHHSHMPKSPQKPIVRVFLPNKQRTVVPARCGVTVRDSLKKALMMRGLIPECCAVYRIQDGEKKPIGWDTDISWLTGEELHVEVLENVPL ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||| ..........MPKSPQKPIVRVFLPNKQRTVVPARCGVTVRDSLKKALMMRGLIPECCAVYRIQDGEKKPIGWDTDISWLTGEELHVEVLENVPL
Dihedral angle restraints
--140-----150-------160-------170-------180-------190-------200-------210-------220-------230-- MGHHHHHHSHMPKSPQKPIVRVFLPNKQRTVVPARCGVTVRDSLKKALMMRGLIPECCAVYRIQDGEKKPIGWDTDISWLTGEELHVEVLENVPL ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||| ...............QKPIVRVFLPNKQRTVVPARCGVTVRDSLKKALMMRGLIPECCAVYRIQDGEKKPIGWDTDISWLTGEELHVEVL --140-----150-------160-------170-------180-------190-------200-------210-------220-------