NMR solution structure of the aromatic mutant H43F H67F cytochrome b5
AEQSDKDVKY YTLEEIQKHK DSKSTWVILH HKVYDLTKFL EEFPGGEEVL REQAGGDATE NFEDVGFSTD ARELSKKYII GELHPDDRSK IAKPSETL
Polymer type: polypeptide(L)
Total | 1H | 13C | 15N | |
---|---|---|---|---|
All | 70.6 % (818 of 1158) | 69.1 % (418 of 605) | 69.1 % (313 of 453) | 87.0 % (87 of 100) |
Backbone | 90.4 % (526 of 582) | 92.0 % (183 of 199) | 89.6 % (258 of 288) | 89.5 % (85 of 95) |
Sidechain | 56.0 % (374 of 668) | 57.9 % (235 of 406) | 53.3 % (137 of 257) | 40.0 % (2 of 5) |
Aromatic | 38.0 % (38 of 100) | 74.0 % (37 of 50) | 0.0 % (0 of 49) | 100.0 % (1 of 1) |
Methyl | 72.3 % (68 of 94) | 85.1 % (40 of 47) | 59.6 % (28 of 47) |
1. Cytochrome b5
AEQSDKDVKY YTLEEIQKHK DSKSTWVILH HKVYDLTKFL EEFPGGEEVL REQAGGDATE NFEDVGFSTD ARELSKKYII GELHPDDRSK IAKPSETLSolvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 283 K, pH 7, Details 0.5 mM [U-99% 15N] FFcytb5, 90% H2O/10% D2O
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
1 | FFcytb5 | [U-99% 15N] | protein | 0.5 mM |
2 | H2O | natural abundance | solvent | 90 % |
3 | D2O | [U-2H] | solvent | 10 % |
4 | potassium phosphate | natural abundance | buffer | 20 mM |
5 | sodium chloride | natural abundance | salt | 20 mM |
6 | Proline+Arginine+Glutamate | natural abundance | salt | 20 mM |
Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 283 K, pH 7, Details 0.5 mM [U-13C; U-15N] FFcytb5, 90% H2O/10% D2O
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
7 | FFcytb5 | [U-13C; U-15N] | protein | 0.5 mM |
8 | H2O | natural abundance | solvent | 90 % |
9 | D2O | [U-2H] | solvent | 10 % |
10 | potassium phosphate | natural abundance | buffer | 20 mM |
11 | sodium chloride | natural abundance | salt | 20 mM |
12 | Proline+Arginine+Glutamate | natural abundance | salt | 20 mM |
Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 283 K, pH 7, Details 0.5 mM [U-13C; U-15N; U-2H] FFcytb5, 90% H2O/10% D2O
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
13 | FFcytb5 | [U-13C; U-15N; U-2H] | protein | 0.5 mM |
14 | H2O | natural abundance | solvent | 90 % |
15 | D2O | [U-2H] | solvent | 10 % |
16 | potassium phosphate | natural abundance | buffer | 20 mM |
17 | sodium chloride | natural abundance | salt | 20 mM |
18 | Proline+Arginine+Glutamate | natural abundance | salt | 20 mM |
Atom type | Mol. common name | Atom group | Value | Ref. method | Ref. type | Shift ratio |
---|---|---|---|---|---|---|
13C | DSS | methyl protons | 0.0 ppm | internal | indirect | 0.2514495 |
1H | DSS | methyl protons | 0.0 ppm | internal | direct | 1.0 |
15N | DSS | methyl protons | 0.0 ppm | internal | indirect | 0.1013291 |
Atom type | Mol. common name | Atom group | Value | Ref. method | Ref. type | Shift ratio |
---|---|---|---|---|---|---|
13C | DSS | methyl protons | 0.0 ppm | internal | indirect | 0.2514495 |
1H | DSS | methyl protons | 0.0 ppm | internal | direct | 1.0 |
15N | DSS | methyl protons | 0.0 ppm | internal | indirect | 0.1013291 |
Atom type | Mol. common name | Atom group | Value | Ref. method | Ref. type | Shift ratio |
---|---|---|---|---|---|---|
13C | DSS | methyl protons | 0.0 ppm | internal | indirect | 0.2514495 |
1H | DSS | methyl protons | 0.0 ppm | internal | direct | 1.0 |
15N | DSS | methyl protons | 0.0 ppm | internal | indirect | 0.1013291 |
Atom type | Mol. common name | Atom group | Value | Ref. method | Ref. type | Shift ratio |
---|---|---|---|---|---|---|
13C | DSS | methyl protons | 0.0 ppm | internal | indirect | 0.2514495 |
1H | DSS | methyl protons | 0.0 ppm | internal | direct | 1.0 |
15N | DSS | methyl protons | 0.0 ppm | internal | indirect | 0.1013291 |
Varian Uniform NMR System - 600 MHz
State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 283 K, pH 7, Details 0.5 mM [U-99% 15N] FFcytb5, 90% H2O/10% D2O
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
1 | FFcytb5 | [U-99% 15N] | protein | 0.5 mM |
2 | H2O | natural abundance | solvent | 90 % |
3 | D2O | [U-2H] | solvent | 10 % |
4 | potassium phosphate | natural abundance | buffer | 20 mM |
5 | sodium chloride | natural abundance | salt | 20 mM |
6 | Proline+Arginine+Glutamate | natural abundance | salt | 20 mM |
Varian Uniform NMR System - 600 MHz
State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 283 K, pH 7, Details 0.5 mM [U-13C; U-15N] FFcytb5, 90% H2O/10% D2O
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
7 | FFcytb5 | [U-13C; U-15N] | protein | 0.5 mM |
8 | H2O | natural abundance | solvent | 90 % |
9 | D2O | [U-2H] | solvent | 10 % |
10 | potassium phosphate | natural abundance | buffer | 20 mM |
11 | sodium chloride | natural abundance | salt | 20 mM |
12 | Proline+Arginine+Glutamate | natural abundance | salt | 20 mM |
Varian Uniform NMR System - 600 MHz
State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 283 K, pH 7, Details 0.5 mM [U-13C; U-15N] FFcytb5, 90% H2O/10% D2O
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
7 | FFcytb5 | [U-13C; U-15N] | protein | 0.5 mM |
8 | H2O | natural abundance | solvent | 90 % |
9 | D2O | [U-2H] | solvent | 10 % |
10 | potassium phosphate | natural abundance | buffer | 20 mM |
11 | sodium chloride | natural abundance | salt | 20 mM |
12 | Proline+Arginine+Glutamate | natural abundance | salt | 20 mM |
Varian Uniform NMR System - 600 MHz
State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 283 K, pH 7, Details 0.5 mM [U-13C; U-15N] FFcytb5, 90% H2O/10% D2O
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
7 | FFcytb5 | [U-13C; U-15N] | protein | 0.5 mM |
8 | H2O | natural abundance | solvent | 90 % |
9 | D2O | [U-2H] | solvent | 10 % |
10 | potassium phosphate | natural abundance | buffer | 20 mM |
11 | sodium chloride | natural abundance | salt | 20 mM |
12 | Proline+Arginine+Glutamate | natural abundance | salt | 20 mM |
Bruker Avance - 700 MHz
State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 283 K, pH 7, Details 0.5 mM [U-13C; U-15N] FFcytb5, 90% H2O/10% D2O
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
7 | FFcytb5 | [U-13C; U-15N] | protein | 0.5 mM |
8 | H2O | natural abundance | solvent | 90 % |
9 | D2O | [U-2H] | solvent | 10 % |
10 | potassium phosphate | natural abundance | buffer | 20 mM |
11 | sodium chloride | natural abundance | salt | 20 mM |
12 | Proline+Arginine+Glutamate | natural abundance | salt | 20 mM |
Varian Uniform NMR System - 600 MHz
State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 283 K, pH 7, Details 0.5 mM [U-13C; U-15N] FFcytb5, 90% H2O/10% D2O
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
7 | FFcytb5 | [U-13C; U-15N] | protein | 0.5 mM |
8 | H2O | natural abundance | solvent | 90 % |
9 | D2O | [U-2H] | solvent | 10 % |
10 | potassium phosphate | natural abundance | buffer | 20 mM |
11 | sodium chloride | natural abundance | salt | 20 mM |
12 | Proline+Arginine+Glutamate | natural abundance | salt | 20 mM |
Bruker Avance - 700 MHz
State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 283 K, pH 7, Details 0.5 mM [U-99% 15N] FFcytb5, 90% H2O/10% D2O
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
1 | FFcytb5 | [U-99% 15N] | protein | 0.5 mM |
2 | H2O | natural abundance | solvent | 90 % |
3 | D2O | [U-2H] | solvent | 10 % |
4 | potassium phosphate | natural abundance | buffer | 20 mM |
5 | sodium chloride | natural abundance | salt | 20 mM |
6 | Proline+Arginine+Glutamate | natural abundance | salt | 20 mM |
Bruker Avance - 700 MHz
State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 283 K, pH 7, Details 0.5 mM [U-13C; U-15N; U-2H] FFcytb5, 90% H2O/10% D2O
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
13 | FFcytb5 | [U-13C; U-15N; U-2H] | protein | 0.5 mM |
14 | H2O | natural abundance | solvent | 90 % |
15 | D2O | [U-2H] | solvent | 10 % |
16 | potassium phosphate | natural abundance | buffer | 20 mM |
17 | sodium chloride | natural abundance | salt | 20 mM |
18 | Proline+Arginine+Glutamate | natural abundance | salt | 20 mM |
Varian Uniform NMR System - 600 MHz
State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 283 K, pH 7, Details 0.5 mM [U-99% 15N] FFcytb5, 90% H2O/10% D2O
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
1 | FFcytb5 | [U-99% 15N] | protein | 0.5 mM |
2 | H2O | natural abundance | solvent | 90 % |
3 | D2O | [U-2H] | solvent | 10 % |
4 | potassium phosphate | natural abundance | buffer | 20 mM |
5 | sodium chloride | natural abundance | salt | 20 mM |
6 | Proline+Arginine+Glutamate | natural abundance | salt | 20 mM |
Varian Uniform NMR System - 600 MHz
State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 283 K, pH 7, Details 0.5 mM [U-13C; U-15N] FFcytb5, 90% H2O/10% D2O
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
7 | FFcytb5 | [U-13C; U-15N] | protein | 0.5 mM |
8 | H2O | natural abundance | solvent | 90 % |
9 | D2O | [U-2H] | solvent | 10 % |
10 | potassium phosphate | natural abundance | buffer | 20 mM |
11 | sodium chloride | natural abundance | salt | 20 mM |
12 | Proline+Arginine+Glutamate | natural abundance | salt | 20 mM |
Properties
Input source #1: NMR data (NEF) - Assigned chemical shifts, Distance restraints, Dihedral angle restraints | combined_36071_5xee.nef |
Input source #2: Coordindates | 5xee.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-------- AEQSDKDVKYYTLEEIQKHKDSKSTWVILHHKVYDLTKFLEEFPGGEEVLREQAGGDATENFEDVGFSTDARELSKKYIIGELHPDDRSKIAKPSETL |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||| AEQSDKDVKYYTLEEIQKHKDSKSTWVILHHKVYDLTKFLEEFPGGEEVLREQAGGDATENFEDVGFSTDARELSKKYIIGELHPDDRSKIAKPSETL
Chain assignments
Entity_assembly_ID (NMR) | Auth_asym_ID (model) | Length | Unmapped | Conflict | Sequence coverage (%) |
---|---|---|---|---|---|
A | A | 98 | 0 | 0 | 100.0 |
Content subtype: combined_36071_5xee.nef
Assigned chemical shifts
--------10--------20--------30--------40--------50--------60--------70--------80--------90-------- AEQSDKDVKYYTLEEIQKHKDSKSTWVILHHKVYDLTKFLEEFPGGEEVLREQAGGDATENFEDVGFSTDARELSKKYIIGELHPDDRSKIAKPSETL |||| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||| ..QSDK.VKYYTLEEIQKHKDSKSTWVILHHKVYDLTKFLEEFPGGEEVLREQAGGDATENFEDVGFSTDARELSKKYIIGELHPDDRSKIAKPSETL
Comp_index_ID | Comp_ID | Atom_ID | CS value (ppm) |
---|---|---|---|
4 | SER | HG | 4.928 |
12 | THR | HG1 | 1.126 |
19 | HIS | HD1 | 6.926 |
22 | SER | HG | 6.87 |
23 | LYS | HZ1 | 7.52 |
23 | LYS | HZ2 | 7.52 |
23 | LYS | HZ3 | 7.52 |
25 | THR | HG1 | 6.675 |
30 | HIS | HD1 | 6.826 |
30 | HIS | HE2 | 7.767 |
31 | HIS | HD1 | 7.294 |
37 | THR | HG1 | 7.056 |
76 | LYS | HZ1 | 6.86 |
76 | LYS | HZ2 | 6.86 |
76 | LYS | HZ3 | 6.86 |
90 | LYS | HZ1 | 7.126 |
90 | LYS | HZ2 | 7.126 |
90 | LYS | HZ3 | 7.126 |
Atom group | # of target shifts | # of assigned shifts | Completeness (%) |
---|---|---|---|
1H chemical shifts | 605 | 403 | 66.6 |
13C chemical shifts | 453 | 302 | 66.7 |
15N chemical shifts | 103 | 84 | 81.6 |
Atom group | # of target shifts | # of assigned shifts | Completeness (%) |
---|---|---|---|
1H chemical shifts | 199 | 179 | 89.9 |
13C chemical shifts | 196 | 174 | 88.8 |
15N chemical shifts | 95 | 83 | 87.4 |
Atom group | # of target shifts | # of assigned shifts | Completeness (%) |
---|---|---|---|
1H chemical shifts | 406 | 224 | 55.2 |
13C chemical shifts | 257 | 128 | 49.8 |
15N chemical shifts | 8 | 1 | 12.5 |
Atom group | # of target shifts | # of assigned shifts | Completeness (%) |
---|---|---|---|
1H chemical shifts | 47 | 38 | 80.9 |
13C chemical shifts | 47 | 25 | 53.2 |
Atom group | # of target shifts | # of assigned shifts | Completeness (%) |
---|---|---|---|
1H chemical shifts | 50 | 37 | 74.0 |
13C chemical shifts | 49 | 0 | 0.0 |
15N chemical shifts | 1 | 1 | 100.0 |
Distance restraints
--------10--------20--------30--------40--------50--------60--------70--------80--------90-------- AEQSDKDVKYYTLEEIQKHKDSKSTWVILHHKVYDLTKFLEEFPGGEEVLREQAGGDATENFEDVGFSTDARELSKKYIIGELHPDDRSKIAKPSETL |||| ||||||||| ||||||||||||||||||||||||||||||||||| ||||||||||||| |||||| |||||||||||||||||| |||| ..QSDK.VKYYTLEEI.KHKDSKSTWVILHHKVYDLTKFLEEFPGGEEVLRE..GGDATENFEDVGF.TDAREL.KKYIIGELHPDDRSKIAK.SETL
--------10--------20--------30--------40--------50--------60--------70--------80--------90-------- AEQSDKDVKYYTLEEIQKHKDSKSTWVILHHKVYDLTKFLEEFPGGEEVLREQAGGDATENFEDVGFSTDARELSKKYIIGELHPDDRSKIAKPSETL | ||| |||| | ||| |||||||||||||||| || |||||| || || ||| ||| | |||||||||| | ||| | ..Q.....KYY.LEEI..H.DSK.TWVILHHKVYDLTKFL.EF..GEEVLR.....DA..NF..VGF..DAR.L.KKYIIGELHP..R.KIA.P --------10--------20--------30--------40--------50--------60--------70--------80--------90----
Dihedral angle restraints
--------10--------20--------30--------40--------50--------60--------70--------80--------90-------- AEQSDKDVKYYTLEEIQKHKDSKSTWVILHHKVYDLTKFLEEFPGGEEVLREQAGGDATENFEDVGFSTDARELSKKYIIGELHPDDRSKIAKPSETL ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||| .EQSDKDVKYYTLEEIQKHKDSKSTWVILHHKVYDLTKFLEEFPGGEEVLREQAGGDATENFEDVGFSTDARELSKKYIIGELHPDDRSKIAKPSETL