3D simutaneous 13C-aromatic,13C-aliphatic,15N edited 1H-1H NOESY
1
8
1
2D 1H-13C HSQC aromatic
1
9
1
3D (H)CCH-COSY aliphatic
1
10
1
3D (H)CCH-COSY aromatic
1
11
1
3D (H)CCH-TOCSY
1
12
2
2D 1H-13C HSQC
1
13
1
2D long-range 1H-15N HSQC for histidine
1
14
1
3DHN(CA)CO
-
試料調製
詳細
Solution-ID
内容
溶媒系
1
0.8 mM [U-100% 13C; U-100% 15N] PgR37A-1, 20 mM MES-2, 100 mM sodium chloride-3, 5 mM calcium chloride-4, 0.02 % sodium azide-5, 50 uM DSS-6, 95% H2O/5% D2O
95% H2O/5% D2O
2
0.8 mM [U-5% 13C; U-100% 15N] PgR37A-7, 20 mM MES-8, 100 mM sodium chloride-9, 5 mM calcium chloride-10, 0.02 % sodium azide-11, 50 uM DSS-12, 95% H2O/5% D2O
95% H2O/5% D2O
試料
濃度 (mg/ml)
構成要素
Isotopic labeling
Solution-ID
0.8mM
PgR37A-1
[U-100% 13C; U-100% 15N]
1
20mM
MES-2
1
100mM
sodium chloride-3
1
5mM
calcium chloride-4
1
0.02 %
sodium azide-5
1
50uM
DSS-6
1
0.8mM
PgR37A-7
[U-5% 13C; U-100% 15N]
2
20mM
MES-8
2
100mM
sodium chloride-9
2
5mM
calcium chloride-10
2
0.02 %
sodium azide-11
2
50uM
DSS-12
2
試料状態
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
Bruker Avance
Bruker
AVANCE
900
1
Varian INOVA
Varian
INOVA
750
2
-
解析
NMR software
名称
バージョン
開発者
分類
CNS
1.2
Brunger, Adams, Clore, Gros, NilgesandRead
精密化
CNS
1.2
Brunger, Adams, Clore, Gros, NilgesandRead
構造決定
CNS
1.2
Brunger, Adams, Clore, Gros, NilgesandRead
geometryoptimization
CYANA
3
Guntert, MumenthalerandWuthrich
構造決定
AutoStructure
2.2.1
Huang, Tejero, PowersandMontelione
構造決定
AutoStructure
2.2.1
Huang, Tejero, PowersandMontelione
structurevalidation
AutoAssign
2.3
Zimmerman, Moseley, KulikowskiandMontelione
データ解析
AutoAssign
2.3
Zimmerman, Moseley, KulikowskiandMontelione
chemicalshiftassignment
CARA
1.8.4
KellerandWuthrich
データ解析
CARA
1.8.4
KellerandWuthrich
peakpicking
CARA
1.8.4
KellerandWuthrich
chemicalshiftassignment
TopSpin
2.1
BrukerBiospin
collection
TopSpin
2.1
BrukerBiospin
解析
VnmrJ
2.1B
Varian
collection
PROSA
6.4
Guntert
解析
TALOS+
1.2009.0721.18
Cornilescu, DelaglioandBax
データ解析
精密化
手法: simulated annealing / ソフトェア番号: 1 詳細: Structure determination was performed iteratively with CYANA v2.1 using NOE-based constraints and PHI and PSI dihedral angle constraints from TALOS+. The 20 conformers out of 100 with the ...詳細: Structure determination was performed iteratively with CYANA v2.1 using NOE-based constraints and PHI and PSI dihedral angle constraints from TALOS+. 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 and upper limit constraints relaxed by 5%
NMR constraints
NOE constraints total: 4039 / NOE intraresidue total count: 637 / NOE long range total count: 1331 / NOE medium range total count: 1134 / NOE sequential total count: 937 / Protein chi angle constraints total count: 0 / Protein other angle constraints total count: 0 / Protein phi angle constraints total count: 77 / Protein psi angle constraints total count: 77
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