1 mM [U-100% 15N] Protein, 20 mM sodium phosphate, 50 mM sodium chloride, 0.1 % sodium azide, 1 mM AEBSF protease inhibitor, 1 mM DTT, 95% H2O/5% D2O
95% H2O/5% D2O
2
1 mM [U-100% 13C; U-100% 15N] Protein, 20 mM sodium phosphate, 50 mM sodium chloride, 0.1 % sodium azide, 1 mM AEBSF protease inhibitor, 1 mM DTT, 95% H2O/5% D2O
95% H2O/5% D2O
3
1 mM [U-100% 13C; U-100% 15N] Protein, 20 mM sodium phosphate, 50 mM sodium chloride, 0.1 % sodium azide, 1 mM AEBSF protease inhibitor, 1 mM DTT, 100% D2O
100% D2O
試料
濃度 (mg/ml)
構成要素
Isotopic labeling
Solution-ID
1mM
Protein-1
[U-100% 15N]
1
20mM
sodium phosphate-2
1
50mM
sodium chloride-3
1
0.1 %
sodium azide-4
1
1mM
AEBSF protease inhibitor-5
1
1mM
DTT-6
1
1mM
Protein-7
[U-100% 13C; U-100% 15N]
2
20mM
sodium phosphate-8
2
50mM
sodium chloride-9
2
0.1 %
sodium azide-10
2
1mM
AEBSF protease inhibitor-11
2
1mM
DTT-12
2
1mM
Protein-13
[U-100% 13C; U-100% 15N]
3
20mM
sodium phosphate-14
3
50mM
sodium chloride-15
3
0.1 %
sodium azide-16
3
1mM
AEBSF protease inhibitor-17
3
1mM
DTT-18
3
試料状態
イオン強度: 50 / pH: 6.5 / 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
Varian INOVA
Varian
INOVA
600
1
Bruker Avance
Bruker
AVANCE
700
2
Bruker Avance
Bruker
AVANCE
800
3
-
解析
NMR software
名称
開発者
分類
NMRPipe
Delaglio, Grzesiek, Vuister, Zhu, PfeiferandBax
解析
CYANA
Guntert, MumenthalerandWuthrich
構造決定
CNS
Brunger, Adams, Clore, Gros, NilgesandRead
精密化
CARA
KellerandWuthrich
chemicalshiftassignment
精密化
手法: molecular dynamics / ソフトェア番号: 1
NMR constraints
NOE constraints total: 2352 / Hydrogen bond constraints total count: 73 / Protein chi angle constraints total count: 17 / Protein phi angle constraints total count: 137 / Protein psi angle constraints total count: 135
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: target function / 計算したコンフォーマーの数: 500 / 登録したコンフォーマーの数: 10