手法: 溶液NMR 詳細: NMR solution structure of the isoform B of bitistatin, a desintegrin isolated from the venom of the viper Bitis arietans
NMR実験
Conditions-ID
Experiment-ID
Solution-ID
タイプ
1
1
1
2D 1H-1H NOESY
1
2
1
2D 1H-1H TOCSY
1
3
1
2D 1H-13C HSQC
1
4
2
2D 1H-15N HSQC
1
5
2
3D 1H-15N NOESY
1
6
2
3D 1H-15N TOCSY
-
試料調製
詳細
Solution-ID
内容
溶媒系
1
1 mM Bitistatin A, 18 mM H2O, 1 mM [U-99% 2H] D2O, 50 mM sodium phosphate, 90% H2O/10% D2O
90% H2O/10% D2O
2
1 mM [U-100% 15N] Bitistatin_B, 18 mM H2O, 1 mM [U-99% 2H] D2O, 50 mM sodium phosphate, 90% H2O/10% D2O
90% H2O/10% D2O
試料
濃度 (mg/ml)
構成要素
Isotopic labeling
Solution-ID
1mM
Bitistatin A-1
1
18mM
H2O-2
1
1mM
D2O-3
[U-99% 2H]
1
50mM
sodium phosphate-4
1
1mM
Bitistatin_B-5
[U-100% 15N]
2
18mM
H2O-6
2
1mM
D2O-7
[U-99% 2H]
2
50mM
sodium phosphate-8
2
試料状態
イオン強度: 50 / pH: 5 / 圧: ambient / 温度: 300 K
-
NMR測定
NMRスペクトロメーター
タイプ: Bruker Avance / 製造業者: Bruker / モデル: AVANCE / 磁場強度: 600 MHz
-
解析
NMR software
名称
開発者
分類
TopSpin
BrukerBiospin
collection
TopSpin
BrukerBiospin
解析
Sparky
Goddard
chemicalshiftassignment
Sparky
Goddard
データ解析
Sparky
Goddard
peakpicking
TALOS
Cornilescu, DelaglioandBax
データ解析
CNSSOLVE
Brunger, Adams, Clore, Gros, NilgesandRead
構造決定
CNSSOLVE
Brunger, Adams, Clore, Gros, NilgesandRead
精密化
精密化
手法: simulated annealing / ソフトェア番号: 1
NMR constraints
NOE constraints total: 1615 / NOE intraresidue total count: 204 / NOE long range total count: 575 / NOE medium range total count: 303 / NOE sequential total count: 479 / Protein phi angle constraints total count: 41 / Protein psi angle constraints total count: 41
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: 43