0.4 mM [U-100% 13C; U-100% 15N] HR7907F, 0.02 % sodium azide, 10 mM DTT, 5 mM calcium chloride, 100 mM sodium chloride, 1 x Proteinase Inhibitors, 20 mM MES, 50 uM DSS, 90% H2O/10% D2O
90% H2O/10% D2O
2
0.3 mM [5% 13C; U-100% 15N] HR7907F, 0.02 % sodium azide, 10 mM DTT, 5 mM calcium chloride, 100 mM sodium chloride, 1 x Proteinase Inhibitors, 20 mM MES, 50 uM DSS, 90% H2O/10% D2O
90% H2O/10% D2O
試料
濃度 (mg/ml)
構成要素
Isotopic labeling
Solution-ID
0.4mM
HR7907F-1
[U-100% 13C; U-100% 15N]
1
0.02 %
sodium azide-2
1
10mM
DTT-3
1
5mM
calcium chloride-4
1
100mM
sodium chloride-5
1
1 %
Proteinase Inhibitors-6
1
20mM
MES-7
1
50uM
DSS-8
1
0.3mM
HR7907F-9
[5% 13C; U-100% 15N]
2
0.02 %
sodium azide-10
2
10mM
DTT-11
2
5mM
calcium chloride-12
2
100mM
sodium chloride-13
2
1 %
Proteinase Inhibitors-14
2
20mM
MES-15
2
50uM
DSS-16
2
試料状態
pH: 6.5 / 圧: 1 atm / 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
Varian INOVA
Varian
INOVA
750
2
-
解析
NMR software
名称
バージョン
開発者
分類
CARA
1.8.4
KellerandWuthrich
peakpicking
CARA
1.8.4
KellerandWuthrich
chemicalshiftassignment
CYANA
3
Guntert, MumenthalerandWuthrich
精密化
CYANA
3
Guntert, MumenthalerandWuthrich
geometryoptimization
CYANA
3
Guntert, MumenthalerandWuthrich
構造決定
AS-DP
1
Huang, Tejero, PowersandMontelione
データ解析
AS-DP
1
Huang, Tejero, PowersandMontelione
精密化
AutoAssign
2.3.0
Zimmerman, Moseley, KulikowskiandMontelione
データ解析
AutoAssign
2.3.0
Zimmerman, Moseley, KulikowskiandMontelione
chemicalshiftassignment
XEASY
Bartelsetal.
データ解析
VnmrJ
Varian
collection
TALOS+
Shen, Cornilescu, DelaglioandBax
geometryoptimization
PSVS
Bhattacharya, Montelione
structurevalidation
精密化
手法: simulated annealing / ソフトェア番号: 1 詳細: STRUCTURE DETERMINATION WAS PERFORMED BY RUNNING CYANA AND ASDP IN PARALLEL USING NOE-BASED CONSTRAINTS AND PHI AND PSI DIHEDRAL ANGLE CONSTRAINTS FROM TALOS+. CONSENSUS PEAK ASSIGNMENTS WERE ...詳細: STRUCTURE DETERMINATION WAS PERFORMED BY RUNNING CYANA AND ASDP IN PARALLEL USING NOE-BASED CONSTRAINTS AND PHI AND PSI DIHEDRAL ANGLE CONSTRAINTS FROM TALOS+. CONSENSUS PEAK ASSIGNMENTS WERE SELECTED AND USED IN ITERATIVE REFINEMENT WITH CYANA. THE 20 CONFORMERS OUT OF 100 WITH THE LOWEST TARGET FUNCTION WERE FURTHER REFINED BY SIMULATED ANNEALING IN EXPLICIT WATER BATH USING THE PROGRAM CNS WITH PARAM19 FORCE FIELD NMR ENSEMBLE INFORMATION
NMR constraints
NOE constraints total: 875 / NOE intraresidue total count: 278 / NOE long range total count: 176 / NOE medium range total count: 217 / NOE sequential total count: 204 / Protein chi angle constraints total count: 0 / Protein other angle constraints total count: 0 / Protein phi angle constraints total count: 36 / Protein psi angle constraints total count: 36
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: 100 / 登録したコンフォーマーの数: 20