RESIDUES 46-67 OF CHAIN E ARE REPLACED WITH A SINGLE SER.
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実験情報
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実験
実験
手法: 溶液NMR
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
3D 1H-15N NOESY
1
4
1
3D 1H-13C NOESY
1
5
1
3D 13C filtered(F1)-13C edited NOESY
1
6
3
2D 1H-15N HSQC
1
7
3
2D 1H-13C HSQC
1
8
3
3D 1H-13C NOESY
1
9
3
3D 1H-15N NOESY
1
10
3
3D 13C filtered(F1)-133D 13C filtered(F1)-13C edited NOESY
1
11
2
3D HNCA
1
12
2
3D HN(CA)CB
1
13
3
3D (H)CCH-TOCSY
1
14
1
3D (H)CCH-TOCSY
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試料調製
詳細
Solution-ID
内容
溶媒系
1
300 uM [U-100% 13C; U-100% 15N] 15N, 13C -NusG-CTD-1, 300 uM NusE-2, 300 uM NusB-3, 90% H2O/10% D2O
90% H2O/10% D2O
2
300 uM [U-100% 13C; U-100% 15N; U-80% 2H] 15N, 13C, 2H-NusE-4, 300 uM NusB-5, 300 uM NusG-CTD-6, 90% H2O/10% D2O
90% H2O/10% D2O
3
0.3 mM NusG-CTD-7, 0.3 mM [U-100% 13C; U-100% 15N] 15N, 13C NusE-8, 0.3 mM NusB-9, 90% H2O/10% D2O
90% H2O/10% D2O
試料
濃度 (mg/ml)
構成要素
Isotopic labeling
Solution-ID
300uM
15N, 13C -NusG-CTD-1
[U-100% 13C; U-100% 15N]
1
300uM
NusE-2
1
300uM
NusB-3
1
300uM
15N, 13C, 2H-NusE-4
[U-100% 13C; U-100% 15N; U-80% 2H]
2
300uM
NusB-5
2
300uM
NusG-CTD-6
2
0.3mM
NusG-CTD-7
3
0.3mM
15N, 13C NusE-8
[U-100% 13C; U-100% 15N]
3
0.3mM
NusB-9
3
試料状態
イオン強度: 50 / pH: 7.5 / 温度: 310 K
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NMR測定
NMRスペクトロメーター
タイプ
製造業者
モデル
磁場強度 (MHz)
Spectrometer-ID
Bruker Avance
Bruker
AVANCE
700
1
Bruker Avance
Bruker
AVANCE
800
2
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解析
NMR software
名称
開発者
分類
XwinNMR
BrukerBiospin
collection
NMRView
Johnson, OneMoonScientific
chemicalshiftassignment
NMRView
Johnson, OneMoonScientific
peakpicking
X-PLOR NIH
Schwieters, Kuszewski, TjandraandClore
構造決定
X-PLOR NIH
Schwieters, Kuszewski, TjandraandClore
精密化
精密化
手法: simulated annealing / ソフトェア番号: 1 詳細: Structure was calculated using rigid body docking with experimental derived intermolecular distance restraints. The structures of unbound NusG-CTD (pdb 2JVV) and unbound NusE:NusB (pdb 3D3B) were used.
代表構造
選択基準: lowest energy
NMRアンサンブル
コンフォーマー選択の基準: structures with the lowest energy 計算したコンフォーマーの数: 20 / 登録したコンフォーマーの数: 18