- PDB-2k1h: Solution NMR structure of SeR13 from Staphylococcus epidermidis. ... -
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基本情報
登録情報
データベース: PDB / ID: 2k1h
タイトル
Solution NMR structure of SeR13 from Staphylococcus epidermidis. Northeast Structural Genomics Consortium target SeR13
要素
Uncharacterized protein SeR13
キーワード
STRUCTURAL GENOMICS / UNKNOWN FUNCTION / PSI-2 / Protein Structure Initiative / Northeast Structural Genomics Consortium / NESG
機能・相同性
機能・相同性情報
Scaffold protein Nfu/NifU, N-terminal domain / Scaffold protein Nfu/NifU, N-terminal / Scaffold protein Nfu/NifU, N-terminal domain superfamily / Scaffold protein Nfu/NifU N terminal / Scaffold protein Nfu/NifU N terminal / Ribosomal Protein S8; Chain: A, domain 1 / 2-Layer Sandwich / Alpha Beta 類似検索 - ドメイン・相同性
Uncharacterized protein / Uncharacterized protein 類似検索 - 構成要素
生物種
Staphylococcus epidermidis (表皮ブドウ球菌)
手法
溶液NMR / simulated annealing
Model details
The oligomeric states of the SeR13 were verified by pulsed-field-gradient diffusion, dynamic light ...The oligomeric states of the SeR13 were verified by pulsed-field-gradient diffusion, dynamic light scattering, sedimentation equilibrium, and concentration dependent studies. The results indicated that the SeR13 self-associates weakly at protein concentration below 1 mM. Using the residual dipolar couplings as well as chemical shift perturbations on dilution, we constructed dimeric models for SeR13. Possible inter-subunit NOEs were identified based on these models. In cases where corresponding NOEs were observed, they were eliminated as constraints and the protein structure recalculated. The absence of structural variation and the failure to observed other predicted inter-subunit NOEs validated use of all NOEs as intra-subunit constraints.
手法: 溶液NMR 詳細: The oligomeric states of the SeR13 were verified by pulsed-field-gradient diffusion, dynamic light scattering, sedimentation equilibrium, and concentration dependent studies. The results ...詳細: The oligomeric states of the SeR13 were verified by pulsed-field-gradient diffusion, dynamic light scattering, sedimentation equilibrium, and concentration dependent studies. The results indicated that the SeR13 self-associates weakly at protein concentration below 1 mM. Using the residual dipolar couplings as well as chemical shift perturbations on dilution, we constructed dimeric models for SeR13. Possible inter-subunit NOEs were identified based on these models. In cases where corresponding NOEs were observed, they were eliminated as constraints and the protein structure recalculated. The absence of structural variation and the failure to observed other predicted inter-subunit NOEs validated use of all NOEs as intra-subunit constraints.
NMR実験
Conditions-ID
Experiment-ID
Solution-ID
タイプ
1
1
1
2D 1H-15N HSQC
1
2
1
2D 1H-13C HSQC
1
3
1
3DCBCA(CO)NH
1
4
1
3DC(CO)NH
1
5
1
3D HNCO
1
6
1
3D HN(CA)CB
1
7
1
3DH(CCO)NH
1
8
1
3D (H)CCH-TOCSY
1
9
1
3D 1H-15N NOESY
1
10
1
3D 1H-15N TOCSY
1
11
1
3D 1H-13C NOESY
1
12
2
2D 1H-15N HSQC-TROSY
1
13
3
2D 1H-15N HSQC-TROSY
-
試料調製
詳細
Solution-ID
内容
溶媒系
1
1.07 mM [U-100% 13C; U-100% 15N] SeR13, 0.2 % sodium azide, 100 mM DTT, 5 mM CaCl2, 100 mM sodium chloride, 20 mM MES, 95% H2O/5% D2O
95% H2O/5% D2O
2
0.86 mM [U-100% 13C; U-100% 15N] SeR13, 0.2 % sodium azide, 100 mM DTT, 5 mM CaCl2, 100 mM sodium chloride, 20 mM MES, 7 % Negative Charged Poly Acrylamide Compressed Gel, 95% H2O/5% D2O
95% H2O/5% D2O
3
0.60 mM [U-100% 13C; U-100% 15N] SeR13, 0.2 % sodium azide, 100 mM DTT, 5 mM CaCl2, 100 mM sodium chloride, 20 mM MES, 7.5 % DMPS/DHPC Mixture, 95% H2O/5% D2O