手法: 溶液NMR 詳細: Second RNA Recognition Motif Domain of hnRNP L bound to ACACAC RNA
NMR実験
Conditions-ID
Experiment-ID
Solution-ID
タイプ
1
1
1
3D 1H-15N NOESY
1
2
2
3D 1H-13C NOESY aliphatic
1
3
2
3D 1H-13C NOESY aromatic
1
4
3
2D 1H-1H NOESY
1
5
3
2D 1H-1H TOCSY
1
6
4
3D F3-filtered-F2-edited NOESY
1
7
4
2D F2-filtered NOESY
-
試料調製
詳細
Solution-ID
内容
溶媒系
1
2 mM [U-100% 15N] protein_1, 2 mM RNA (5'-R(*AP*CP*AP*CP*AP*C)-3'), 60 mM sodium chloride, 40 mM sodium phosphate, 1 mM DTT, 90% H2O/10% D2O
90% H2O/10% D2O
2
2 mM [U-100% 13C; U-100% 15N] protein_1, 2 mM RNA (5'-R(*AP*CP*AP*CP*AP*C)-3'), 60 mM sodium chloride, 40 mM sodium phosphate, 1 mM DTT, 90% H2O/10% D2O
90% H2O/10% D2O
3
2 mM [U-100% 15N] protein_1, 2 mM protein_1, 60 mM sodium chloride, 40 mM sodium phosphate, 1 mM DTT, 100% D2O
100% D2O
4
2 mM [U-100% 13C; U-100% 15N] protein_1, 2 mM RNA (5'-R(*AP*CP*AP*CP*AP*C)-3'), 60 mM sodium chloride, 40 mM sodium phosphate, 1 mM DTT, 100% D2O
100% D2O
試料
濃度 (mg/ml)
構成要素
Isotopic labeling
Solution-ID
2mM
entity_1-1
[U-100% 15N]
1
2mM
RNA (5'-R(*AP*CP*AP*CP*AP*C)-3')-2
1
60mM
sodium chloride-3
1
40mM
sodium phosphate-4
1
1mM
DTT-5
1
2mM
entity_1-6
[U-100% 13C; U-100% 15N]
2
2mM
RNA (5'-R(*AP*CP*AP*CP*AP*C)-3')-7
2
60mM
sodium chloride-8
2
40mM
sodium phosphate-9
2
1mM
DTT-10
2
2mM
entity_1-11
[U-100% 15N]
3
2mM
entity_1-12
3
60mM
sodium chloride-13
3
40mM
sodium phosphate-14
3
1mM
DTT-15
3
2mM
entity_1-16
[U-100% 13C; U-100% 15N]
4
2mM
RNA (5'-R(*AP*CP*AP*CP*AP*C)-3')-17
4
60mM
sodium chloride-18
4
40mM
sodium phosphate-19
4
1mM
DTT-20
4
試料状態
イオン強度: 0.1 / pH: 6.5 / 圧: ambient / 温度: 310.15 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
900
1
Bruker Avance
Bruker
AVANCE
700
2
-
解析
NMR software
名称
開発者
分類
TopSpin
BrukerBiospin
collection
TopSpin
BrukerBiospin
解析
Sparky
Goddard
chemicalshiftassignment
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
代表構造
選択基準: closest to the average
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: 250 / 登録したコンフォーマーの数: 20