手法: 溶液NMR 詳細: The solution structure of the C-terminal domain of Prp24, containing an RRM with flanking alpha helices.
NMR実験
Conditions-ID
Experiment-ID
Solution-ID
タイプ
1
1
1
2D 1H-15N HSQC
1
2
1
3DCBCA(CO)NH
1
3
1
3D 1H-15N NOESY
1
4
1
3D HN(CA)CB
1
5
1
3D HNCO
1
6
1
3DHBHA(CO)NH
1
7
1
3DC(CO)NH
1
8
1
3DH(CCO)NH
1
9
2
2D 1H-13C HSQC aliphatic
1
10
2
2D 1H-13C HSQC aromatic
1
11
4
2D 1H-1H NOESY
1
12
2
3D (H)CCH-TOCSY
1
13
2
3D 1H-13C NOESY aliphatic
1
14
2
3D 1H-13C NOESY aromatic
1
15
3
2D 1H-13C HSQC aliphatic
2
16
1
2D-NHSQC-IPAP
2
17
5
2D-NHSQC-IPAP
2
18
1
3D-Jmodulated-CHSQC
2
19
5
3D-Jmodulated-CHSQC
-
試料調製
詳細
Solution-ID
内容
溶媒系
1
600 uM [U-99% 13C; U-99% 15N] L4W, 18 mM potassium phosphate pH 6, 45 mM potassium chloride, 0.9 mM DTT, 90% H2O/10% D2O
90% H2O/10% D2O
2
600 uM [U-99% 13C; U-99% 15N] L4W, 18 mM [U-99% 2H] potassium phosphate pH 6, 45 mM [U-99% 2H] potassium chloride, 100% D2O
100% D2O
3
600 uM [U-99% 13C; U-99% 15N] L4W, 18 mM potassium phosphate pH 6, 45 mM potassium chloride, 1 uM DSS, 90% H2O/10% D2O
90% H2O/10% D2O
4
600 uM L4W, 18 mM [U-99% 2H] potassium phosphate pH 6, 45 mM [U-99% 2H] potassium chloride, 100% D2O
100% D2O
5
600 uM [U-99% 13C; U-99% 15N] L4W, 18 mM potassium phosphate pH 6, 45 mM potassium chloride, 0.9 mM DTT, 6.5 % DMPC/DHPC q=3, 0.67 mg/mL CTAB, 90% H2O/10% D2O
90% H2O/10% D2O
試料
濃度 (mg/ml)
構成要素
Isotopic labeling
Solution-ID
600uM
L4W-1
[U-99% 13C; U-99% 15N]
1
18mM
potassium phosphate pH 6-2
1
45mM
potassium chloride-3
1
0.9mM
DTT-4
1
600uM
L4W-5
[U-99% 13C; U-99% 15N]
2
18mM
potassium phosphate pH 6-6
[U-99% 2H]
2
45mM
potassium chloride-7
[U-99% 2H]
2
600uM
L4W-8
[U-99% 13C; U-99% 15N]
3
18mM
potassium phosphate pH 6-9
3
45mM
potassium chloride-10
3
1uM
DSS-11
3
600uM
L4W-12
4
18mM
potassium phosphate pH 6-13
[U-99% 2H]
4
45mM
potassium chloride-14
[U-99% 2H]
4
600uM
L4W-15
[U-99% 13C; U-99% 15N]
5
18mM
potassium phosphate pH 6-16
5
45mM
potassium chloride-17
5
0.9mM
DTT-18
5
6.5 %
DMPC/DHPC q=3-19
5
0.67mg/mL
CTAB-20
5
試料状態
Conditions-ID
pH
圧 (kPa)
温度 (K)
1
6
ambient
298K
2
6
ambient
304K
-
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
Varian Uniform NMR System
Varian
UniformNMRSystem
600
1
Bruker DMX
Bruker
DMX
750
2
Varian Uniform NMR System
Varian
UniformNMRSystem
900
3
-
解析
NMR software
名称
バージョン
開発者
分類
VnmrJ
Varian
collection
XwinNMR
BrukerBiospin
collection
NMRDraw
Delaglio, Grzesiek, Vuister, Zhu, PfeiferandBax
解析
Sparky
Goddard
chemicalshiftassignment
Sparky
Goddard
データ解析
Sparky
Goddard
peakpicking
TALOS
Cornilescu, DelaglioandBax
データ解析
HADDOCK
2
AlexandreBonvin
精密化
HADDOCK
2
AlexandreBonvin
データ解析
UNIO
8
TorstenHerrmann
構造決定
NMR Structure Tools
LawrenceJClos
データ解析
精密化
手法: simulated annealing / ソフトェア番号: 1
NMR constraints
NOE constraints total: 1379 / NOE intraresidue total count: 460 / NOE long range total count: 284 / NOE medium range total count: 272 / NOE sequential total count: 363 / Hydrogen bond constraints total count: 53
代表構造
選択基準: lowest average pairwise rmsd
NMRアンサンブル
コンフォーマー選択の基準: all calculated structures submitted 計算したコンフォーマーの数: 20 / 登録したコンフォーマーの数: 20 / Maximum lower distance constraint violation: 0 Å / Maximum upper distance constraint violation: 0.333 Å