150 mM sodium chloride-1, 10 mM HEPES-2, 3 mM TCEP-3, 7 % [U-100% 2H] D2O-4, 5 mM Octyl Glucoside-5, 93 % H2O-6, 99 % [U-99% 15N] N15 labeled PAS-7, 93% H2O/7% D2O
93% H2O/7% D2O
2
150 mM sodium chloride-8, 10 mM HEPES-9, 3 mM TCEP-10, 100 % [U-100% 2H] D2O-11, 5 mM Octyl Glucoside-12, 99 % [U-99% 13C; U-99% 15N] N15/C13 labeled PAS-13, 100% D2O
100% D2O
試料
濃度 (mg/ml)
構成要素
Isotopic labeling
Solution-ID
150mM
sodium chloride-1
1
10mM
HEPES-2
1
3mM
TCEP-3
1
7 %
D2O-4
[U-100% 2H]
1
5mM
Octyl Glucoside-5
1
93 %
H2O-6
1
99 %
N15 labeled PAS-7
[U-99% 15N]
1
150mM
sodium chloride-8
2
10mM
HEPES-9
2
3mM
TCEP-10
2
100 %
D2O-11
[U-100% 2H]
2
5mM
Octyl Glucoside-12
2
99 %
N15/C13 labeled PAS-13
[U-99% 13C; U-99% 15N]
2
試料状態
イオン強度: 150mM / pH: 6.9 / 圧: AMBIENT / 温度: 300 K
-
NMR測定
NMRスペクトロメーター
タイプ: BRUKER Avance II / 製造業者: Bruker / モデル: AVANCE II / 磁場強度: 900 MHz
-
解析
NMR software
名称
バージョン
開発者
分類
CYANA
2.1
Guntert, MumenthalerandWuthrich
geometryoptimization
CYANA
2.1
Guntert, MumenthalerandWuthrich
精密化
Amber
10
Case, Darden, Cheatham, III, Simmerling, Wang, Duke, Luo, andKollm
精密化
XEASY
Bartelsetal.
chemicalshiftassignment
XEASY
Bartelsetal.
データ解析
XEASY
Bartelsetal.
peakpicking
TALOS
Cornilescu, DelaglioandBax
dihedralanglescalculation
TopSpin
2
BrukerBiospin
collection
TopSpin
2
BrukerBiospin
解析
精密化
手法: DGSA-distance geometry simulated annealing, Generalised Born Energy Minimization, GENERALISED BORN ENERGY MINIMIZATION WITH NMR RESTRAINTS ソフトェア番号: 1 詳細: CYANA 2.1, AMBER 10 with NMR restraints, FORMATION 3 ENERGY 0.0000
NMR constraints
NOE constraints total: 2634 / NOE long range total count: 877 / NOE medium range total count: 498 / NOE sequential total count: 1259 / Hydrogen bond constraints total count: 96
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: structures with the lowest energy 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: 20 / 代表コンフォーマー: 1