手法: 溶液NMR 詳細: High resolution NMR solution structure of a complex of HIV-2 TAR RNA and a synthetic tripeptide in a 1:2 stoichiometry
NMR実験
Conditions-ID
Experiment-ID
Solution-ID
タイプ
2
1
1
2D 1H-15N HSQC
1
2
2
2D 1H-13C HSQC
1
3
3
2D 1H-1H TOCSY
1
4
3
2D 1H-1H NOESY
1
5
4
2D 1H-1H NOESY
1
6
2
3D (H)CCH-TOCSY
1
7
2
3D (H)CCH-COSY
2
8
1
3D 1H-15N NOESY
1
9
2
3D 1H-13C NOESY
1
10
2
2DH(C)N
1
11
2
2D 1H-15N HSQC (2J)
1
12
2
2D 1H-13C HSQC
2
13
1
2D HNN-COSY
1
14
2
2D H5NN-COSY
1
15
2
3D forward-directed HCC-TOCSY-CCH E.COSY
2
16
1
2D 1H-15N CPMG-NOESY
-
試料調製
詳細
Solution-ID
内容
溶媒系
1
0.65 mM [U-100% 13C; U-100% 15N] UUCG-TAR-1, 5.2 mM Pyrimidinylpeptide-2, 50 mM potassium chloride-3, 25 mM potassium phosphate-4, 90% H2O/10% D2O
90% H2O/10% D2O
2
0.65 mM [U-100% 13C; U-100% 15N] UUCG-TAR-5, 5.2 mM Pyrimidinylpeptide-6, 50 mM potassium chloride-7, 25 mM potassium phosphate-8, 100% D2O
100% D2O
3
0.65 mM UUCG-TAR-9, 5.2 mM Pyrimidinylpeptide-10, 50 mM potassium chloride-11, 25 mM potassium phosphate-12, 90% H2O / 10% D2O
90% H2O/10% D2O
4
0.65 mM UUCG-TAR-13, 5.2 mM Pyrimidinylpeptide-14, 50 mM potassium chloride-15, 25 mM potassium phosphate-16, 100% D2O
100% D2O
5
0.2 mM [U-100% 13C; U-100% 15N] UUCG-TAR-17, 0.8 mM Pyrimidinylpeptide-18, 50 mM potassium chloride-19, 25 mM potassium phosphate-20, 16 mg/mL Pf1 phage-21, 100% D2O
100% D2O
試料
濃度 (mg/ml)
構成要素
Isotopic labeling
Solution-ID
0.65mM
UUCG-TAR-1
[U-100% 13C; U-100% 15N]
1
5.2mM
Pyrimidinylpeptide-2
1
50mM
potassium chloride-3
1
25mM
potassium phosphate-4
1
0.65mM
UUCG-TAR-5
[U-100% 13C; U-100% 15N]
2
5.2mM
Pyrimidinylpeptide-6
2
50mM
potassium chloride-7
2
25mM
potassium phosphate-8
2
0.65mM
UUCG-TAR-9
3
5.2mM
Pyrimidinylpeptide-10
3
50mM
potassium chloride-11
3
25mM
potassium phosphate-12
3
0.65mM
UUCG-TAR-13
4
5.2mM
Pyrimidinylpeptide-14
4
50mM
potassium chloride-15
4
25mM
potassium phosphate-16
4
0.2mM
UUCG-TAR-17
[U-100% 13C; U-100% 15N]
5
0.8mM
Pyrimidinylpeptide-18
5
50mM
potassium chloride-19
5
25mM
potassium phosphate-20
5
16mg/mL
Pf1 phage-21
5
試料状態
Conditions-ID
イオン強度
pH
圧 (kPa)
温度 (K)
1
75
6.2
ambient
298K
2
75
6.2
ambient
283K
-
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
800
2
Bruker Avance
Bruker
AVANCE
700
3
Bruker Avance
Bruker
AVANCE
600
4
Bruker DRX
Bruker
DRX
600
5
Bruker Avance
Bruker
AVANCE
400
6
-
解析
NMR software
名称
バージョン
開発者
分類
TopSpin
2.1
BrukerBiospin
collection
TopSpin
2.1
BrukerBiospin
解析
TopSpin
2.1
BrukerBiospin
データ解析
Sparky
3.114
Goddard
chemicalshiftassignment
Sparky
3.114
Goddard
データ解析
Sparky
3.114
Goddard
peakpicking
CNS
1.1
Brunger, Adams, Clore, Gros, NilgesandRead
構造決定
ARIA
1.2
Linge, O'DonoghueandNilges
精密化
HADDOCK
2.1
Dominguez, BoelensandBonvin
精密化
精密化
手法: simulated annealing / ソフトェア番号: 1
NMR constraints
NA alpha-angle constraints total count: 22 / NA beta-angle constraints total count: 4 / NA chi-angle constraints total count: 27 / NA epsilon-angle constraints total count: 5 / NA other-angle constraints total count: 22 / NA sugar pucker constraints total count: 95 / NOE constraints total: 567 / Hydrogen bond constraints total count: 60