0.7 mM [U-100% 13C; U-100% 15N; U-80% 2H] protein, 50 mM HEPES, 50 mM sodium chloride, 2 mM DTT, 1 mM EDTA, 0.1 % sodium azide, 95% H2O/5% D2O
95% H2O/5% D2O
2
0.7 mM [U-100% 15N] protein, 50 mM HEPES, 50 mM sodium chloride, 2 mM DTT, 1 mM EDTA, 0.1 % sodium azide, 95% H2O/5% D2O
95% H2O/5% D2O
3
0.7 mM [U-100% 13C] protein, 50 mM HEPES, 50 mM sodium chloride, 2 mM DTT, 1 mM EDTA, 0.1 % sodium azide, 100% D2O
100% D2O
試料
濃度 (mg/ml)
構成要素
Isotopic labeling
Solution-ID
0.7mM
protein-1
[U-100% 13C; U-100% 15N; U-80% 2H]
1
50mM
HEPES-2
1
50mM
sodium chloride-3
1
2mM
DTT-4
1
1mM
EDTA-5
1
0.1 %
sodium azide-6
1
0.7mM
protein-7
[U-100% 15N]
2
50mM
HEPES-8
2
50mM
sodium chloride-9
2
2mM
DTT-10
2
1mM
EDTA-11
2
0.1 %
sodium azide-12
2
0.7mM
protein-13
[U-100% 13C]
3
50mM
HEPES-14
3
50mM
sodium chloride-15
3
2mM
DTT-16
3
1mM
EDTA-17
3
0.1 %
sodium azide-18
3
試料状態
イオン強度: 50 / pH: 6.7 / 圧: ambient / 温度: 298 K
-
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
850
1
Bruker Avance
Bruker
AVANCE
900
2
Bruker Avance
Bruker
AVANCE
700
3
Varian INOVA
Varian
INOVA
800
4
-
解析
NMR software
名称
開発者
分類
TopSpin
BrukerBiospin
collection
NMRPipe
Delaglio, Grzesiek, Vuister, Zhu, PfeiferandBax
解析
XEASY
Bartelsetal.
chemicalshiftassignment
XEASY
Bartelsetal.
peakpicking
XEASY
Bartelsetal.
データ解析
TALOS
Cornilescu, DelaglioandBax
データ解析
X-PLOR NIH
Schwieters, Kuszewski, TjandraandClore
構造決定
X-PLOR NIH
Schwieters, Kuszewski, TjandraandClore
精密化
精密化
手法: simulated annealing / ソフトェア番号: 1
NMR constraints
NOE constraints total: 1242 / NOE intraresidue total count: 576 / NOE long range total count: 247 / NOE medium range total count: 241 / NOE sequential total count: 178 / Protein phi angle constraints total count: 91 / Protein psi angle constraints total count: 91
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: 50 / 登録したコンフォーマーの数: 10 / Maximum lower distance constraint violation: 1.8 Å / Maximum upper distance constraint violation: 5.5 Å