back calculated data agree with experimental NOESY spectrum,structures with acceptable covalent geometry,structures with the least restraint violations, structures with the lowest energy
代表モデル
モデル #1
minimized average structure
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要素
#1: RNA鎖
PHOSPHOROTHIOATESUBSTITUTEDPHAGEMS2RNABINDINGSITE
分子量: 6809.375 Da / 分子数: 1 / 由来タイプ: 合成 詳細: Transcription by T7 RNA ploymerase using 5'-a-thio-ATP
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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 NOESY
1
2
1
3D 15N-separated NOESY
2
3
2
3D 15N-separated NOESY
2
4
2
DQF-COSY
2
5
2
31P-1H HETCOR
2
6
2
2D 13C-filtered NOESY
NMR実験の詳細
Text: The structure was determined using standard 2D homonuclear and 2D/3D heteronuclear techniques. Filtered experiments used to identify inter-residue NOEs involving unlabeled A residues
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試料調製
詳細
Solution-ID
内容
溶媒系
1
2.0 mM replicase operator RNA U-15N,13C at U, C, and G, unlabeled A; phosphorothioate 5' to each adenine;; 10 mM KPi buffer; 10 mM NaCl; 0.2 mM EDTA
90% H2O/10% D2O
2
2.0 mM replicase operator RNA U-15N,13C at U, C, and G, unlabeled A; phosphorothioate 5' to each adenine;; 10 mM KPi buffer; 10 mM NaCl; 0.2 mM EDTA
100% D2O
試料状態
イオン強度: ~50mM / pH: 6.8 / 圧: ambient / 温度: 298 K
結晶化
*PLUS
手法: other / 詳細: NMR
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NMR測定
NMRスペクトロメーター
タイプ: Bruker AMX / 製造業者: Bruker / モデル: AMX / 磁場強度: 500 MHz
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解析
NMR software
名称
バージョン
開発者
分類
UXNMR
1994
Spectraspin
collection
Felix
1998
Biosym-MSI
解析
X-PLOR
3.851
Brunger
構造決定
X-PLOR
3.851
Brunger
精密化
精密化
手法: simulated annealing, molecular dynamics refinement / ソフトェア番号: 1 詳細: structures were calculated using 313 conformationally restrictive NOE distance constraints (i.e., intra-residue sugar-sugar NOEs are not included), 20 H-bond base pair constraints, 30 ...詳細: structures were calculated using 313 conformationally restrictive NOE distance constraints (i.e., intra-residue sugar-sugar NOEs are not included), 20 H-bond base pair constraints, 30 backbone dihedral restraints, and dihedral restraints to restrict sugar pucker conformation. Parameter file was modified to incorporate bond-length, bond-angle, charge, van der Waal's radius for the Rp phosphoryl sulfur atoms located 5' to each adenine in the sequence. [Jaroszewski, JW., et al., Anti-Cancer Drug Design ( 1992) 7, 253; Florian, J. et al., J. Am. Chem. Soc. (1998) 120, 7959].
代表構造
選択基準: minimized average structure
NMRアンサンブル
コンフォーマー選択の基準: back calculated data agree with experimental NOESY spectrum,structures with acceptable covalent geometry,structures with the least restraint violations, ...コンフォーマー選択の基準: back calculated data agree with experimental NOESY spectrum,structures with acceptable covalent geometry,structures with the least restraint violations, structures with the lowest energy 計算したコンフォーマーの数: 30 / 登録したコンフォーマーの数: 10