1.5 mM [U-99% 13C; U-99% 15N] protein, 10 mM sodium phosphate, 200 mM sodium chloride, 10 mM DTT, 0.01 % sodium azide, 1 mM benzamidine, 93% H2O/7% D2O
93% H2O/7% D2O
2
1.5 mM [U-7% 13C; U-99% 15N] protein, 10 mM sodium phosphate, 200 mM sodium chloride, 10 mM DTT, 0.01 % sodium azide, 1 mM benzamidine, 93% H2O/7% D2O
93% H2O/7% D2O
試料
濃度 (mg/ml)
構成要素
Isotopic labeling
Solution-ID
1.5mM
protein
[U-99% 13C; U-99% 15N]
1
10mM
sodiumphosphate
1
200mM
sodiumchloride
1
10mM
DTT
1
0.01 %
sodiumazide
1
1mM
benzamidine
1
1.5mM
protein
[U-7% 13C; U-99% 15N]
2
10mM
sodiumphosphate
2
200mM
sodiumchloride
2
10mM
DTT
2
0.01 %
sodiumazide
2
1mM
benzamidine
2
試料状態
イオン強度: 200 / pH: 6.5 / 圧: ambient / 温度: 293 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
Varian INOVA
Varian
INOVA
750
2
Varian INOVA
Varian
INOVA
800
3
-
解析
NMR software
名称
バージョン
開発者
分類
CYANA
3
Guntert, P. etal.
構造決定
Sparky
3.11
Goddard, T. etal.
データ解析
Sparky
3.11
Goddard, T. etal.
peakpicking
Felix
2007
AccelrysSoftwareInc.
解析
CNS
1.1
Brunger, A. etal.
精密化
CNS
1.1
Brunger, A. etal.
構造決定
PSVS
1.3
Bhattacharya, A. etal.
データ解析
精密化
手法: torsion angle dynamics, molecular dynamics / ソフトェア番号: 1 詳細: Structure determination was performed iteratively using CYANA (Automated NOESY Assignments). A total of 20 structures out of 100 with lowest target function from the final CYANA calculation ...詳細: Structure determination was performed iteratively using CYANA (Automated NOESY Assignments). A total of 20 structures out of 100 with lowest target function from the final CYANA calculation were taken and refined by restrained molecular dynamics/energy minimization in explicit water (CNS) after adding 15% to the upper boundary limit. The final set of structures contained no dihedral violations greater than 1 degree and only 4 distance violations total that were greater than 0.05 Angstrom.
NMR constraints
NOE constraints total: 1619 / NOE intraresidue total count: 425 / NOE long range total count: 319 / NOE medium range total count: 399 / NOE sequential total count: 476 / Hydrogen bond constraints total count: 60 / Protein chi angle constraints total count: 0 / Protein other angle constraints total count: 0 / Protein phi angle constraints total count: 63 / Protein psi angle constraints total count: 63
代表構造
選択基準: closest to the average
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: 100 / 登録したコンフォーマーの数: 20