0.2 mM [U-13C; U-15N], segmentally labeled RNA (1-16 LABELED), 1.2 mM [U-100% 15N; U-80% 2H] RsmE, 0.03 mM sodium chloride, 0.05 mM potassium phosphate, 95% H2O/5% D2O
95% H2O/5% D2O
2
0.2 mM [U-13C; U-15N], segmentally labeled RNA (17-40 LABELED), 1.2 mM [U-100% 15N; U-80% 2H] RsmE, 0.03 mM sodium chloride, 0.05 mM potassium phosphate, 95% H2O/5% D2O
95% H2O/5% D2O
3
0.2 mM [U-13C; U-15N], segmentally labeled RNA (41-58 LABELED), 1.2 mM [U-100% 15N; U-80% 2H] RsmE, 0.03 mM sodium chloride, 0.05 mM potassium phosphate, 95% H2O/5% D2O
95% H2O/5% D2O
4
0.2 mM [U-13C; U-15N], segmentally labeled RNA (59-72 LABELED), 1.2 mM [U-100% 15N; U-80% 2H] RsmE, 0.03 mM sodium chloride, 0.05 mM potassium phosphate, 95% H2O/5% D2O
95% H2O/5% D2O
5
0.2 mM [U-13C; U-15N], segmentally labeled RNA (36-44 LABELED), 1.2 mM [U-100% 15N; U-80% 2H] RsmE, 0.03 mM sodium chloride, 0.05 mM potassium phosphate, 95% H2O/5% D2O
95% H2O/5% D2O
6
0.2 mM RNA (72-MER), 1.2 mM [U-100% 15N; U-80% 2H] RsmE, 0.03 mM sodium chloride, 0.05 mM potassium phosphate, 95% H2O/5% D2O
95% H2O/5% D2O
試料
濃度 (mg/ml)
構成要素
Isotopic labeling
Solution-ID
0.2mM
RNA (72-MER)-1
[U-13C; U-15N], segmentally labeled
1
1.2mM
RsmE-2
[U-100% 15N; U-80% 2H]
1
0.03mM
sodium chloride-3
1
0.05mM
potassium phosphate-4
1
0.2mM
RNA (72-MER)-5
[U-13C; U-15N], segmentally labeled
2
1.2mM
RsmE-6
[U-100% 15N; U-80% 2H]
2
0.03mM
sodium chloride-7
2
0.05mM
potassium phosphate-8
2
0.2mM
RNA (72-MER)-9
[U-13C; U-15N], segmentally labeled
3
1.2mM
RsmE-10
[U-100% 15N; U-80% 2H]
3
0.03mM
sodium chloride-11
3
0.05mM
potassium phosphate-12
3
0.2mM
RNA (72-MER)-13
[U-13C; U-15N], segmentally labeled
4
1.2mM
RsmE-14
[U-100% 15N; U-80% 2H]
4
0.03mM
sodium chloride-15
4
0.05mM
potassium phosphate-16
4
0.2mM
RNA (72-MER)-17
[U-13C; U-15N], segmentally labeled
5
1.2mM
RsmE-18
[U-100% 15N; U-80% 2H]
5
0.03mM
sodium chloride-19
5
0.05mM
potassium phosphate-20
5
0.2mM
RNA (72-MER)-21
6
1.2mM
RsmE-22
[U-100% 15N; U-80% 2H]
6
0.03mM
sodium chloride-23
6
0.05mM
potassium phosphate-24
6
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
CYANA
Guntert, MumenthalerandWuthrich
構造決定
Amber
Case, Darden, Cheatham, III, Simmerling, Wang, Duke, LuoandKollman
精密化
TopSpin
BrukerBiospin
解析
Sparky
Goddard
データ解析
精密化
手法: simulated annealing / ソフトェア番号: 1
NMR constraints
NA sugar pucker constraints total count: 29 / NOE constraints total: 7441 / Protein phi angle constraints total count: 288 / Protein psi angle constraints total count: 292
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: 2500 / 登録したコンフォーマーの数: 20 / Maximum torsion angle constraint violation: 1.22 ° / Maximum upper distance constraint violation: 0.3 Å