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Membrane protein complex DsbB-DsbA structure by joint calculations with solid-state NMR and X-ray experimental data
Authors
Tang, M., Sperling, L.J., Berthold, D.A., Schwieters, C.D., Nesbitt, A.E., Nieuwkoop, A.J., Gennis, R.B., Rienstra, C.M.
Assembly
DsbA and DsbB
Entity
1. DsbA (polymer), 188 monomers, 21003.63 Da Detail

AQYEDGKQYT TLEKPVAGAP QVLEFFSFFC PHCYQFEEVL HISDNVKKKL PEGVKMTKYH VNFMGGDLGK DLTQAWAVAM ALGVEDKVTV PLFEGVQKTQ TIRSASDIRD VFINAGIKGE EYDAAWNSFV VKSLVAQQEK AAADVQLRGV PAMFVNGKYQ LNPQGMDTSN MDVFVQQYAD TVKYLSEK


2. DsbB (polymer), 176 monomers, 20089.81 Da Detail

MLRFLNQASQ GRGAWLLMAF TALALELTAL WFQHVMLLKP SVLCIYERVA LFGVLGAALI GAIAPKTPLR YVAMVIWLYS AFRGVQLTYE HTMLQLYPSP FATCDFMVRF PEWLPLDKWV PQVFVASGDC AERQWDFLGL EMPQWLLGIF IAYLIVAVLV VISQPFKAKK RDLFGR


3. ZN (non-polymer), 65.409 Da
4. UQ8 (non-polymer), 727.109 Da
Total weight
41885.96 Da
Max. entity weight
21003.63 Da
Entity Connection
disulfide 2 Detail

IDTypeValue orderAtom ID 1Atom ID 2
1disulfidesing1:CYS30:SG1:CYS33:SG
2disulfidesing2:CYS104:SG2:CYS130:SG

Source organism
Escherichia coli
Exptl. method
NMR
Refine. method
simulated annealing
Data set
assigned_chemical_shifts
Chem. Shift Complete
Sequence coverage: 91.8 %, Completeness: 34.8 %, Completeness (bb): 56.8 % Detail

Polymer type: polypeptide(L)

Total1H13C15N
All34.8 % (1531 of 4394) 4.3 % (96 of 2258)66.4 % (1153 of 1737)70.7 % (282 of 399)
Backbone56.8 % (1208 of 2127)10.1 % (72 of 711)80.3 % (858 of 1069)80.1 % (278 of 347)
Sidechain22.7 % (593 of 2608) 1.6 % (24 of 1547)56.0 % (565 of 1009) 7.7 % (4 of 52)
Aromatic 6.3 % (30 of 480) 0.0 % (0 of 240)12.1 % (28 of 231)22.2 % (2 of 9)
Methyl40.4 % (185 of 458) 0.0 % (0 of 229)80.8 % (185 of 229)

1. DsbA

AQYEDGKQYT TLEKPVAGAP QVLEFFSFFC PHCYQFEEVL HISDNVKKKL PEGVKMTKYH VNFMGGDLGK DLTQAWAVAM ALGVEDKVTV PLFEGVQKTQ TIRSASDIRD VFINAGIKGE EYDAAWNSFV VKSLVAQQEK AAADVQLRGV PAMFVNGKYQ LNPQGMDTSN MDVFVQQYAD TVKYLSEK

2. DsbB

MLRFLNQASQ GRGAWLLMAF TALALELTAL WFQHVMLLKP SVLCIYERVA LFGVLGAALI GAIAPKTPLR YVAMVIWLYS AFRGVQLTYE HTMLQLYPSP FATCDFMVRF PEWLPLDKWV PQVFVASGDC AERQWDFLGL EMPQWLLGIF IAYLIVAVLV VISQPFKAKK RDLFGR

Sample #1

Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 270 K, pH 7.0


#NameIsotope labelingTypeConcentration
1DsbA[U-100% 13C; U-100% 15N]15 mg
2H2Onatural abundance90 %
3D2Onatural abundance10 %
Sample #2

Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 270 K, pH 7.0


#NameIsotope labelingTypeConcentration
4DsbA[2-13C-glycerol; U-15N]10 mg
5H2Onatural abundance90 %
6D2Onatural abundance10 %
Sample #3

Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 270 K, pH 7.0


#NameIsotope labelingTypeConcentration
7DsbA[1,3-13C-glycerol; U-15N]10 mg
8H2Onatural abundance90 %
9D2Onatural abundance10 %
Sample #4

Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 270 K, pH 7.0


#NameIsotope labelingTypeConcentration
10DsbB[U-100% 13C; U-100% 15N]7 mg
11DDMnatural abundance2 mg
12E. coli lipidsnatural abundance7 mg
13H2Onatural abundance90 %
14D2Onatural abundance10 %
Sample #5

Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 270 K, pH 7.0


#NameIsotope labelingTypeConcentration
15DsbB[2-13C-glycerol; U-15N]5 mg
16DDMnatural abundance2 mg
17E. coli lipidsnatural abundance7 mg
18H2Onatural abundance90 %
19D2Onatural abundance10 %
Sample #6

Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 270 K, pH 7.0


#NameIsotope labelingTypeConcentration
20DsbB[1,3-13C-glycerol; U-15N]4 mg
21DDMnatural abundance2 mg
22E. coli lipidsnatural abundance7 mg
23H2Onatural abundance90 %
24D2Onatural abundance10 %

LACS Plot; CA
Referencing offset: -0.14 ppm, Outliers: 1 Detail
LACS Plot; CB
Referencing offset: -0.14 ppm, Outliers: 1 Detail
LACS Plot; CO
Referencing offset: -0.69 ppm, Outliers: 1 Detail
Protein Blocks Logo
Calculated from 10 models in PDB: 2LEG, Strand ID: A, B Detail


Release date
2011-10-11
Citation
High-resolution membrane protein structure by joint calculations with solid-state NMR and X-ray experimental data
Tang, M., Sperling, L.J., Berthold, D.A., Schwieters, C.D., Nesbitt, A.E., Nieuwkoop, A.J., Gennis, R.B., Rienstra, C.M.
J. Biomol. NMR (2011), 51, 227-233, PubMed 21938394 , DOI 10.1007/s10858-011-9565-6 ,
Related entities 2. DsbB, : 1 : 2 : 6 : 66 entities Detail
Interaction partners 2. DsbB, : 1 interactors Detail
Keywords DsbB-DsbA, Joint calculations, Solid-state NMR, X-ray reflections