Solution structure of BCL-xL determined with selective isotope labelling of I,L,V sidechains
MSMAMSQSNR ELVVDFLSYK LSQKGYSWSQ FSDVEENRTE APEGTESEAV KQALREAGDE FELRYRRAFS DLTSQLHITP GTAYQSFEQV VNELFRDGVN WGRIVAFFSF GGALCVESVD KEMQVLVSRI AAWMATYLND HLEPWIQENG GWDTFVELYG NNAAAESRKG QERLEHHHHH H
Polymer type: polypeptide(L)
Total | 1H | 13C | 15N | |
---|---|---|---|---|
All | 49.3 % (1039 of 2106) | 29.4 % (320 of 1089) | 67.4 % (550 of 816) | 84.1 % (169 of 201) |
Backbone | 77.0 % (832 of 1080) | 50.9 % (189 of 371) | 90.2 % (479 of 531) | 92.1 % (164 of 178) |
Sidechain | 30.0 % (358 of 1195) | 18.2 % (131 of 718) | 48.9 % (222 of 454) | 21.7 % (5 of 23) |
Aromatic | 8.7 % (21 of 240) | 9.2 % (11 of 120) | 4.3 % (5 of 115) | 100.0 % (5 of 5) |
Methyl | 78.6 % (132 of 168) | 75.0 % (63 of 84) | 82.1 % (69 of 84) |
1. BCL-xL apo
MSMAMSQSNR ELVVDFLSYK LSQKGYSWSQ FSDVEENRTE APEGTESEAV KQALREAGDE FELRYRRAFS DLTSQLHITP GTAYQSFEQV VNELFRDGVN WGRIVAFFSF GGALCVESVD KEMQVLVSRI AAWMATYLND HLEPWIQENG GWDTFVELYG NNAAAESRKG QERLEHHHHH HSolvent system 92% H2O/8% D2O, Pressure 1 atm, Temperature 273 K, pH 7.0 (±0.05), Details 10 mM Na phosphate 40 mM NaCl 0.1% NaN3 pH 7.0
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
1 | BCL-xL_apo | [U-100% 13C; U-100% 15N; U-95% 2H; Ile,Leu,Val C1H3] | 0.9 mM | |
2 | sodium phosphate | natural abundance | 10 mM | |
3 | sodium chloride | natural abundance | 40 mM | |
4 | NaN3 | natural abundance | 0.1 % | |
5 | H2O | natural abundance | 92 % | |
6 | D2O | natural abundance | 8 % |
Atom type | Mol. common name | Atom group | Value | Ref. method | Ref. type | Shift ratio |
---|---|---|---|---|---|---|
13C | DSS | methyl protons | 0.0 ppm | external | indirect | 0.25144 |
1H | DSS | methyl protons | 0.0 ppm | external | direct | 1.0 |
15N | [15N] nitromethane | nitrogen | 0.0 ppm | external | direct | 1.0 |
Atom type | Mol. common name | Atom group | Value | Ref. method | Ref. type | Shift ratio |
---|---|---|---|---|---|---|
13C | DSS | methyl protons | 0.0 ppm | external | indirect | 0.25144 |
1H | DSS | methyl protons | 0.0 ppm | external | direct | 1.0 |
15N | [15N] nitromethane | nitrogen | 0.0 ppm | external | direct | 1.0 |
Atom type | Mol. common name | Atom group | Value | Ref. method | Ref. type | Shift ratio |
---|---|---|---|---|---|---|
13C | DSS | methyl protons | 0.0 ppm | external | indirect | 0.25144 |
1H | DSS | methyl protons | 0.0 ppm | external | direct | 1.0 |
15N | [15N] nitromethane | nitrogen | 0.0 ppm | external | direct | 1.0 |
Bruker Avance - 800 MHz cryogenically-cooled, triple-resonance single-axis gradient probe
State isotropic, Solvent system 92% H2O/8% D2O, Pressure 1 atm, Temperature 273 K, pH 7.0 (±0.05), Details 10 mM Na phosphate 40 mM NaCl 0.1% NaN3 pH 7.0
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
1 | BCL-xL_apo | [U-100% 13C; U-100% 15N; U-95% 2H; Ile,Leu,Val C1H3] | 0.9 mM | |
2 | sodium phosphate | natural abundance | 10 mM | |
3 | sodium chloride | natural abundance | 40 mM | |
4 | NaN3 | natural abundance | 0.1 % | |
5 | H2O | natural abundance | 92 % | |
6 | D2O | natural abundance | 8 % |
Bruker Avance - 800 MHz cryogenically-cooled, triple-resonance single-axis gradient probe
State isotropic, Solvent system 92% H2O/8% D2O, Pressure 1 atm, Temperature 273 K, pH 7.0 (±0.05), Details 10 mM Na phosphate 40 mM NaCl 0.1% NaN3 pH 7.0
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
1 | BCL-xL_apo | [U-100% 13C; U-100% 15N; U-95% 2H; Ile,Leu,Val C1H3] | 0.9 mM | |
2 | sodium phosphate | natural abundance | 10 mM | |
3 | sodium chloride | natural abundance | 40 mM | |
4 | NaN3 | natural abundance | 0.1 % | |
5 | H2O | natural abundance | 92 % | |
6 | D2O | natural abundance | 8 % |
Bruker Avance - 800 MHz cryogenically-cooled, triple-resonance single-axis gradient probe
State isotropic, Solvent system 92% H2O/8% D2O, Pressure 1 atm, Temperature 273 K, pH 7.0 (±0.05), Details 10 mM Na phosphate 40 mM NaCl 0.1% NaN3 pH 7.0
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
1 | BCL-xL_apo | [U-100% 13C; U-100% 15N; U-95% 2H; Ile,Leu,Val C1H3] | 0.9 mM | |
2 | sodium phosphate | natural abundance | 10 mM | |
3 | sodium chloride | natural abundance | 40 mM | |
4 | NaN3 | natural abundance | 0.1 % | |
5 | H2O | natural abundance | 92 % | |
6 | D2O | natural abundance | 8 % |
Bruker Avance - 800 MHz cryogenically-cooled, triple-resonance single-axis gradient probe
State isotropic, Solvent system 92% H2O/8% D2O, Pressure 1 atm, Temperature 273 K, pH 7.0 (±0.05), Details 10 mM Na phosphate 40 mM NaCl 0.1% NaN3 pH 7.0
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
1 | BCL-xL_apo | [U-100% 13C; U-100% 15N; U-95% 2H; Ile,Leu,Val C1H3] | 0.9 mM | |
2 | sodium phosphate | natural abundance | 10 mM | |
3 | sodium chloride | natural abundance | 40 mM | |
4 | NaN3 | natural abundance | 0.1 % | |
5 | H2O | natural abundance | 92 % | |
6 | D2O | natural abundance | 8 % |
Bruker Avance - 800 MHz cryogenically-cooled, triple-resonance single-axis gradient probe
State isotropic, Solvent system 92% H2O/8% D2O, Pressure 1 atm, Temperature 273 K, pH 7.0 (±0.05), Details 10 mM Na phosphate 40 mM NaCl 0.1% NaN3 pH 7.0
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
1 | BCL-xL_apo | [U-100% 13C; U-100% 15N; U-95% 2H; Ile,Leu,Val C1H3] | 0.9 mM | |
2 | sodium phosphate | natural abundance | 10 mM | |
3 | sodium chloride | natural abundance | 40 mM | |
4 | NaN3 | natural abundance | 0.1 % | |
5 | H2O | natural abundance | 92 % | |
6 | D2O | natural abundance | 8 % |
Bruker Avance - 800 MHz cryogenically-cooled, triple-resonance single-axis gradient probe
State isotropic, Solvent system 92% H2O/8% D2O, Pressure 1 atm, Temperature 273 K, pH 7.0 (±0.05), Details 10 mM Na phosphate 40 mM NaCl 0.1% NaN3 pH 7.0
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
1 | BCL-xL_apo | [U-100% 13C; U-100% 15N; U-95% 2H; Ile,Leu,Val C1H3] | 0.9 mM | |
2 | sodium phosphate | natural abundance | 10 mM | |
3 | sodium chloride | natural abundance | 40 mM | |
4 | NaN3 | natural abundance | 0.1 % | |
5 | H2O | natural abundance | 92 % | |
6 | D2O | natural abundance | 8 % |
Bruker Avance - 800 MHz cryogenically-cooled, triple-resonance single-axis gradient probe
State isotropic, Solvent system 92% H2O/8% D2O, Pressure 1 atm, Temperature 273 K, pH 7.0 (±0.05), Details 10 mM Na phosphate 40 mM NaCl 0.1% NaN3 pH 7.0
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
1 | BCL-xL_apo | [U-100% 13C; U-100% 15N; U-95% 2H; Ile,Leu,Val C1H3] | 0.9 mM | |
2 | sodium phosphate | natural abundance | 10 mM | |
3 | sodium chloride | natural abundance | 40 mM | |
4 | NaN3 | natural abundance | 0.1 % | |
5 | H2O | natural abundance | 92 % | |
6 | D2O | natural abundance | 8 % |
Bruker Avance - 800 MHz cryogenically-cooled, triple-resonance single-axis gradient probe
State isotropic, Solvent system 92% H2O/8% D2O, Pressure 1 atm, Temperature 273 K, pH 7.0 (±0.05), Details 10 mM Na phosphate 40 mM NaCl 0.1% NaN3 pH 7.0
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
1 | BCL-xL_apo | [U-100% 13C; U-100% 15N; U-95% 2H; Ile,Leu,Val C1H3] | 0.9 mM | |
2 | sodium phosphate | natural abundance | 10 mM | |
3 | sodium chloride | natural abundance | 40 mM | |
4 | NaN3 | natural abundance | 0.1 % | |
5 | H2O | natural abundance | 92 % | |
6 | D2O | natural abundance | 8 % |
Bruker Avance - 800 MHz cryogenically-cooled, triple-resonance single-axis gradient probe
State isotropic, Solvent system 92% H2O/8% D2O, Pressure 1 atm, Temperature 273 K, pH 7.0 (±0.05), Details 10 mM Na phosphate 40 mM NaCl 0.1% NaN3 pH 7.0
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
1 | BCL-xL_apo | [U-100% 13C; U-100% 15N; U-95% 2H; Ile,Leu,Val C1H3] | 0.9 mM | |
2 | sodium phosphate | natural abundance | 10 mM | |
3 | sodium chloride | natural abundance | 40 mM | |
4 | NaN3 | natural abundance | 0.1 % | |
5 | H2O | natural abundance | 92 % | |
6 | D2O | natural abundance | 8 % |
Bruker Avance - 800 MHz cryogenically-cooled, triple-resonance single-axis gradient probe
State isotropic, Solvent system 92% H2O/8% D2O, Pressure 1 atm, Temperature 273 K, pH 7.0 (±0.05), Details 10 mM Na phosphate 40 mM NaCl 0.1% NaN3 pH 7.0
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
1 | BCL-xL_apo | [U-100% 13C; U-100% 15N; U-95% 2H; Ile,Leu,Val C1H3] | 0.9 mM | |
2 | sodium phosphate | natural abundance | 10 mM | |
3 | sodium chloride | natural abundance | 40 mM | |
4 | NaN3 | natural abundance | 0.1 % | |
5 | H2O | natural abundance | 92 % | |
6 | D2O | natural abundance | 8 % |
Bruker Avance - 800 MHz cryogenically-cooled, triple-resonance single-axis gradient probe
State isotropic, Solvent system 92% H2O/8% D2O, Pressure 1 atm, Temperature 273 K, pH 7.0 (±0.05), Details 10 mM Na phosphate 40 mM NaCl 0.1% NaN3 pH 7.0
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
1 | BCL-xL_apo | [U-100% 13C; U-100% 15N; U-95% 2H; Ile,Leu,Val C1H3] | 0.9 mM | |
2 | sodium phosphate | natural abundance | 10 mM | |
3 | sodium chloride | natural abundance | 40 mM | |
4 | NaN3 | natural abundance | 0.1 % | |
5 | H2O | natural abundance | 92 % | |
6 | D2O | natural abundance | 8 % |
Properties
Input source #1: NMR data (NEF) - Assigned chemical shifts, Distance restraints, Dihedral angle restraints | combined_18792_2m03.nef |
Input source #2: Coordindates | 2m03.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
--------10--------20--------30--------40--------50--------60--------70--------80--------90-------100 MSMAMSQSNRELVVDFLSYKLSQKGYSWSQFSDVEENRTEAPEGTESEAVKQALREAGDEFELRYRRAFSDLTSQLHITPGTAYQSFEQVVNELFRDGVN |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||| MSMAMSQSNRELVVDFLSYKLSQKGYSWSQFSDVEENRTEAPEGTESEAVKQALREAGDEFELRYRRAFSDLTSQLHITPGTAYQSFEQVVNELFRDGVN -------110-------120-------130-------140-------150-------160-------170-------180- WGRIVAFFSFGGALCVESVDKEMQVLVSRIAAWMATYLNDHLEPWIQENGGWDTFVELYGNNAAAESRKGQERLEHHHHHH ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||| WGRIVAFFSFGGALCVESVDKEMQVLVSRIAAWMATYLNDHLEPWIQENGGWDTFVELYGNNAAAESRKGQERLEHHHHHH
Chain assignments
Entity_assembly_ID (NMR) | Auth_asym_ID (model) | Length | Unmapped | Conflict | Sequence coverage (%) |
---|---|---|---|---|---|
A | A | 181 | 0 | 0 | 100.0 |
Content subtype: combined_18792_2m03.nef
Assigned chemical shifts
--------10--------20--------30--------40--------50--------60--------70--------80--------90-------100 MSMAMSQSNRELVVDFLSYKLSQKGYSWSQFSDVEENRTEAPEGTESEAVKQALREAGDEFELRYRRAFSDLTSQLHITPGTAYQSFEQVVNELFRDGVN |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||| ||| ||||||||||||| ...AMSQSNRELVVDFLSYKLSQKGYSWSQFSDVEENRTEAPEGTESEAVKQALREAGDEFELRYRRAFSDLTSQLHIT....YQS.EQVVNELFRDGVN --------10--------20--------30--------40--------50--------60--------70--------80--------90-------100 -------110-------120-------130-------140-------150-------160-------170-------180- WGRIVAFFSFGGALCVESVDKEMQVLVSRIAAWMATYLNDHLEPWIQENGGWDTFVELYGNNAAAESRKGQERLEHHHHHH |||||||||||||||||||||| |||||||||||||||||||||||||||||||||||||||||||||||||||| WGRIVAFFSFGGALCVESVDKE.QVLVSRIAAWMATYLNDHLEPWIQENGGWDTFVELYGNNAAAESRKGQERLE -------110-------120-------130-------140-------150-------160-------170-----
Comp_index_ID | Comp_ID | Atom_ID | CS value (ppm) |
---|---|---|---|
42 | PRO | N | 135.09 |
144 | PRO | N | 134.541 |
Atom group | # of target shifts | # of assigned shifts | Completeness (%) |
---|---|---|---|
1H chemical shifts | 1089 | 227 | 20.8 |
13C chemical shifts | 816 | 536 | 65.7 |
15N chemical shifts | 212 | 169 | 79.7 |
Atom group | # of target shifts | # of assigned shifts | Completeness (%) |
---|---|---|---|
1H chemical shifts | 371 | 164 | 44.2 |
13C chemical shifts | 362 | 328 | 90.6 |
15N chemical shifts | 178 | 164 | 92.1 |
Atom group | # of target shifts | # of assigned shifts | Completeness (%) |
---|---|---|---|
1H chemical shifts | 718 | 63 | 8.8 |
13C chemical shifts | 454 | 208 | 45.8 |
15N chemical shifts | 34 | 5 | 14.7 |
Atom group | # of target shifts | # of assigned shifts | Completeness (%) |
---|---|---|---|
1H chemical shifts | 89 | 58 | 65.2 |
13C chemical shifts | 89 | 67 | 75.3 |
Atom group | # of target shifts | # of assigned shifts | Completeness (%) |
---|---|---|---|
1H chemical shifts | 120 | 5 | 4.2 |
13C chemical shifts | 115 | 0 | 0.0 |
15N chemical shifts | 5 | 5 | 100.0 |
Distance restraints
--------10--------20--------30--------40--------50--------60--------70--------80--------90-------100 MSMAMSQSNRELVVDFLSYKLSQKGYSWSQFSDVEENRTEAPEGTESEAVKQALREAGDEFELRYRRAFSDLTSQLHITPGTAYQSFEQVVNELFRDGVN |||||||||||||||||||||||||||| ||| |||||||||||||||||||||||||||||||||||| ||| ||||||||||||| .......SNRELVVDFLSYKLSQKGYSWSQFSDVE...TEA.EGTESEAVKQALREAGDEFELRYRRAFSDLTSQLHI.....YQS.EQVVNELFRDGVN --------10--------20--------30--------40--------50--------60--------70--------80--------90-------100 -------110-------120-------130-------140-------150-------160-------170-------180- WGRIVAFFSFGGALCVESVDKEMQVLVSRIAAWMATYLNDHLEPWIQENGGWDTFVELYGNNAAAESRKGQERLEHHHHHH |||||||||||||||||||||| |||||||||||||||||||| ||||||||||||||||||||||| | WGRIVAFFSFGGALCVESVDKE.QVLVSRIAAWMATYLNDHLE.WIQENGGWDTFVELYGNNAAAES......L -------110-------120-------130-------140-------150-------160-------170----
Dihedral angle restraints
--------10--------20--------30--------40--------50--------60--------70--------80--------90-------100 MSMAMSQSNRELVVDFLSYKLSQKGYSWSQFSDVEENRTEAPEGTESEAVKQALREAGDEFELRYRRAFSDLTSQLHITPGTAYQSFEQVVNELFRDGVN ||||||||||||||||||||||||||||||| |||||||||||||||||||||||||||||||| ||||||||||||||| | ..MAMSQSNRELVVDFLSYKLSQKGYSWSQFSD...........TESEAVKQALREAGDEFELRYRRAFSDLTSQL......AYQSFEQVVNELFRD..N --------10--------20--------30--------40--------50--------60--------70--------80--------90-------100 -------110-------120-------130-------140-------150-------160-------170-------180- WGRIVAFFSFGGALCVESVDKEMQVLVSRIAAWMATYLNDHLEPWIQENGGWDTFVELYGNNAAAESRKGQERLEHHHHHH |||||||||||||||||||||||||||||||||||||||||| |||||||||||||||| ||||||| |||| WGRIVAFFSFGGALCVESVDKEMQVLVSRIAAWMATYLNDHL.PWIQENGGWDTFVELY..NAAAESR......EHHH -------110-------120-------130-------140-------150-------160-------170--------