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Solution NMR structure of integral membrane protein DsbB
Authors
Zhou, Y., Cierpicki, T., Flores Jimenez, R.H., Lukasik, S.M., Ellena, J.F., Cafiso, D.S., Kadukura, H., Beckwith, J., Bushweller, J.H.
Assembly
DsbB
Entity
1. DsbB (polymer, Thiol state: all disulfide bound), 183 monomers, 20925.58 Da Detail

MLRFLNQASQ GRGAWLLMAF TALALELTAL WFQHVMLLKP CVLSIYERAA LFGVLGAALI GAIAPKTPLR YVAMVIWLYS AFRGVQLTYE HTMLQLYPSP FATSDFMVRF PEWLPLDKWV PQVFVASGDC AERQWDFLGL EMPQWLLGIF IAYLIVAVLV VISQPFKAKK RDLFGRGHHH HHH


Formula weight
20925.58 Da
Entity Connection
disulfide 1 Detail

IDTypeValue orderAtom ID 1Atom ID 2
1disulfidesing1:CYS41:SG1:CYS130:SG

Source organism
Escherichia coli
Exptl. method
solution NMR
Refine. method
simulated annealing
Data set
assigned_chemical_shifts
Chem. Shift Complete
Sequence coverage: 95.1 %, Completeness: 41.9 %, Completeness (bb): 77.3 % Detail

Polymer type: polypeptide(L)

Total1H13C15N
All41.9 % (945 of 2256)21.4 % (247 of 1152)58.1 % (526 of 905)86.4 % (172 of 199)
Backbone77.3 % (825 of 1067)46.6 % (166 of 356)92.0 % (495 of 538)94.8 % (164 of 173)
Sidechain20.4 % (278 of 1361)10.2 % (81 of 796)35.1 % (189 of 539)30.8 % (8 of 26)
Aromatic18.4 % (54 of 294)19.0 % (28 of 147)13.6 % (19 of 140)100.0 % (7 of 7)
Methyl13.5 % (34 of 252) 6.3 % (8 of 126)20.6 % (26 of 126)

1. Disulfide bond formation protein B

MLRFLNQASQ GRGAWLLMAF TALALELTAL WFQHVMLLKP CVLSIYERAA LFGVLGAALI GAIAPKTPLR YVAMVIWLYS AFRGVQLTYE HTMLQLYPSP FATSDFMVRF PEWLPLDKWV PQVFVASGDC AERQWDFLGL EMPQWLLGIF IAYLIVAVLV VISQPFKAKK RDLFGRGHHH HHH

Sample #1

Solvent system 95% H2O/5% D2O, Pressure 1 atm, Temperature 313 K, pH 6.2


#NameIsotope labelingTypeConcentration
1DsbB[CSSC][U-13C; U-15N; U-2H]1.2 mM
2DPC[U-2H]100 mM
3sodium phosphatenatural abundance25 mM
4potassium chloridenatural abundance50 mM
Sample #2

Solvent system 95% H2O/5% D2O, Pressure 1 atm, Temperature 313 K, pH 6.2


#NameIsotope labelingTypeConcentration
5DsbB[CSSC]I,L,V methyl protonated, [U-13C; U-15N; U-2H]1.5 mM
6DPC[U-2H]100 mM
7sodium phosphatenatural abundance25 mM
8potassium chloridenatural abundance50 mM
Sample #3

Solvent system 95% H2O/5% D2O, Pressure 1 atm, Temperature 313 K, pH 6.2, Details single Cys mutants to incorporate MTSL/dMTSL for PRE measurements


#NameIsotope labelingTypeConcentration
9DsbB[CSSC] single Cys mutants[U-15N; U-2H]0.5 ~ 1.0 mM
10DPCnatural abundance100 mM
11sodium phosphatenatural abundance25 mM
12potassium chloridenatural abundance50 mM

LACS Plot; CA
Referencing offset: 0.46 ppm, Outliers: 1 Detail
LACS Plot; CB
Referencing offset: 0.46 ppm, Outliers: 1 Detail
LACS Plot; CO
Referencing offset: 0.25 ppm, Outliers: 3 Detail
Protein Blocks Logo
Calculated from 20 models in PDB: 2K73, Strand ID: A Detail


Release date
2008-12-11
Citation
NMR solution structure of the integral membrane enzyme DsbB: functional insights into DsbB-catalyzed disulfide bond formation
Zhou, Y., Cierpicki, T., Flores Jimenez, R.H., Lukasik, S.M., Ellena, J.F., Cafiso, D.S., Kadukura, H., Beckwith, J., Bushweller, J.H.
Mol. Cell (2008), 31, 896-908, PubMed 18922471 , DOI 10.1016/j.molcel.2008.08.028 ,
Related entities 1. DsbB, : 1 : 1 : 7 : 53 entities Detail
Interaction partners 1. DsbB, : 1 interactors Detail
Experiments performed 13 experiments Detail
Keywords disulfide bond, DsbB, membrane protein, redox enzyme