1 mM [U-100% 15N] RsmE, 1 mM SL2(RsmZ), 0.05 mM potassium phosphate, 0.03 mM sodium chloride, 95% H2O/5% D2O
95% H2O/5% D2O
2
1 mM [U-100% 15N] RsmE, 1 mM SL2(RsmZ), 0.05 mM potassium phosphate, 0.03 mM sodium chloride, 100% D2O
100% D2O
3
1 mM [U-100% 13C; U-100% 15N] RsmE, 1 mM SL2(RsmZ), 0.05 mM potassium phosphate, 0.03 mM sodium chloride, 95% H2O/5% D2O
95% H2O/5% D2O
4
1 mM [U-100% 13C; U-100% 15N] RsmE, 1 mM SL2(RsmZ), 0.05 mM potassium phosphate, 0.03 mM sodium chloride, 100% D2O
100% D2O
5
1 mM [U-100% 15N] RsmE, 1 mM [U-100% 13C; U-100% 15N]-Ade/Ura RNA SL2(RsmZ), 0.05 mM potassium phosphate, 0.03 mM sodium chloride, 95% H2O/5% D2O
95% H2O/5% D2O
6
1 mM [U-100% 15N] RsmE, 1 mM [U-100% 13C; U-100% 15N]-Ade/Ura RNA SL2(RsmZ), 0.05 mM potassium phosphate, 0.03 mM sodium chloride, 100% D2O
100% D2O
7
1 mM [U-100% 15N] RsmE, 1 mM [U-100% 13C; U-100% 15N]-Cyt/Gua RNA SL2(RsmZ), 0.05 mM potassium phosphate, 0.03 mM sodium chloride, 95% H2O/5% D2O
95% H2O/5% D2O
8
1 mM [U-100% 15N] RsmE, 1 mM [U-100% 13C; U-100% 15N]-Cyt/Gua RNA SL2(RsmZ), 0.05 mM potassium phosphate, 0.03 mM sodium chloride, 100% D2O
100% D2O
試料
濃度 (mg/ml)
構成要素
Isotopic labeling
Solution-ID
1mM
RsmE-1
[U-100% 15N]
1
1mM
SL2(RsmZ)-2
1
0.05mM
potassium phosphate-3
1
0.03mM
sodium chloride-4
1
1mM
RsmE-5
[U-100% 15N]
2
1mM
SL2(RsmZ)-6
2
0.05mM
potassium phosphate-7
2
0.03mM
sodium chloride-8
2
1mM
RsmE-9
[U-100% 13C; U-100% 15N]
3
1mM
SL2(RsmZ)-10
3
0.05mM
potassium phosphate-11
3
0.03mM
sodium chloride-12
3
1mM
RsmE-13
[U-100% 13C; U-100% 15N]
4
1mM
SL2(RsmZ)-14
4
0.05mM
potassium phosphate-15
4
0.03mM
sodium chloride-16
4
1mM
RsmE-17
[U-100% 15N]
5
1mM
SL2(RsmZ)-18
[U-100% 13C; U-100% 15N]-Ade/Ura RNA
5
0.05mM
potassium phosphate-19
5
0.03mM
sodium chloride-20
5
1mM
RsmE-21
[U-100% 15N]
6
1mM
SL2(RsmZ)-22
[U-100% 13C; U-100% 15N]-Ade/Ura RNA
6
0.05mM
potassium phosphate-23
6
0.03mM
sodium chloride-24
6
1mM
RsmE-25
[U-100% 15N]
7
1mM
SL2(RsmZ)-26
[U-100% 13C; U-100% 15N]-Cyt/Gua RNA
7
0.05mM
potassium phosphate-27
7
0.03mM
sodium chloride-28
7
1mM
RsmE-29
[U-100% 15N]
8
1mM
SL2(RsmZ)-30
[U-100% 13C; U-100% 15N]-Cyt/Gua RNA
8
0.05mM
potassium phosphate-31
8
0.03mM
sodium chloride-32
8
試料状態
イオン強度: 0.18 / pH: 7.2 / 圧: ambient / 温度: 313 K
-
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
500
1
Bruker Avance
Bruker
AVANCE
600
2
Bruker Avance
Bruker
AVANCE
700
3
Bruker Avance
Bruker
AVANCE
900
4
-
解析
NMR software
名称
開発者
分類
TopSpin
BrukerBiospin
解析
Sparky
Goddard
chemicalshiftassignment
Sparky
Goddard
データ解析
CYANA
Guntert, MumenthalerandWuthrich
構造決定
Amber
Case, Darden, Cheatham, III, Simmerling, Wang, Duke, Luo, ... andKollman
精密化
精密化
手法: simulated annealing / ソフトェア番号: 1
NMR constraints
NA sugar pucker constraints total count: 12 / NOE constraints total: 2700
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: 999 / 登録したコンフォーマーの数: 20 / Maximum torsion angle constraint violation: 13.7 ° / Maximum upper distance constraint violation: 0.42 Å