Text: Backbone and side chain 1H, 15N and 13C resonances were assigned using standard triple resonance experiments. Distance restraints were derived from 13C- and 15N-edited 3D NOESY experiments. ...Text: Backbone and side chain 1H, 15N and 13C resonances were assigned using standard triple resonance experiments. Distance restraints were derived from 13C- and 15N-edited 3D NOESY experiments. Assignments for the RNA were obtained from 2D isotope-filtered experiments. Intermolecular distance restraints were measured in 3D 13C-edited/filtered experiments. Dihedral angle restraints for the backbone angle phi were derived from 3J(HN,Ha) coupling constants measured in an HNHA-J experiment, additional phi/psi restraints were derived from TALOS. Hydrogen bond restraints for secondary structure elements in the protein were defined from slowly exchanging amide protons, identified after exchange of the H2O buffer to D2O.
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試料調製
詳細
Solution-ID
内容
溶媒系
1
1 mM U-15N,13C SF1 KH-QUA2/unlabeled BPS in 20 mM phosphate buffer NA pH 6.5, 50 mM NaCl, 2 mM DTT
90% H2O/10% D2O
2
1 mM U-15N,13C SF1 KH-QUA2/unlabeled BPS in 20 mM phosphate buffer NA pH 6.5, 50 mM NaCl, 2 mM DTT
100% D2O
3
1 mM U-15N SF1 KH-QUA2/unlabeled BPS in 20 mM phosphate buffer NA pH 6.5, 50 mM NaCl, 2 mM DTT
90% H2O/10% D2O
試料状態
Conditions-ID
イオン強度
pH
圧 (kPa)
温度 (K)
1
50mMNaCl
6.5
ambient
295K
2
50mMNaCl
6.5
ambient
278K
結晶化
*PLUS
手法: その他 / 詳細: NMR
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NMR測定
放射
プロトコル: SINGLE WAVELENGTH / 単色(M)・ラウエ(L): M
放射波長
相対比: 1
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 DRX
Bruker
DRX
500
1
Bruker DRX
Bruker
DRX
600
2
Bruker DRX
Bruker
DRX
800
3
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解析
NMR software
名称
バージョン
開発者
分類
XwinNMR
2.6
Bruker
collection
NMRPipe
1.8
F. Delaglio, S. Grzesiek, G. Vuister, G. Zhu, J. Pfeifer, & A. Bax
解析
XEASY
1.2
Ch. Bartels, T.-H. Xia, M. Billeter, P. Gntert and K. Wthrich
データ解析
ARIA/CNS
1
J. Linge, M. Nilges
構造決定
ARIA/CNS
1
J. Linge, M. Nilges
精密化
精密化
手法: restrained molecular dynamics, simulated annealing / ソフトェア番号: 1 詳細: STRUCTURES WERE CALCULATED WITH A MIXED TORSION AND CARTESIAN ANGLE DYNAMICS PROTOCOL USING ARIA/CNS. A REDUCED RELAXATION MATRIX APPROACH WAS USED FOR THE NOE CALIBRATION. THE STRUCTURES ARE ...詳細: STRUCTURES WERE CALCULATED WITH A MIXED TORSION AND CARTESIAN ANGLE DYNAMICS PROTOCOL USING ARIA/CNS. A REDUCED RELAXATION MATRIX APPROACH WAS USED FOR THE NOE CALIBRATION. THE STRUCTURES ARE CURRENTLY BEING REFINED USING RESIDUAL DIPOLAR COUPLINGS AND WILL BE UPDATED IN THE FUTURE.
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: 10 lowest energy structures consistent with experimental distance and dihedral angle restraints 計算したコンフォーマーの数: 100 / 登録したコンフォーマーの数: 10