Text: The authors state that the structures are arranged by energy, preceeding from lowest to highest, but there is no single representative structure. Binding of Gcn4 residues 101-134 to Gal11 ...Text: The authors state that the structures are arranged by energy, preceeding from lowest to highest, but there is no single representative structure. Binding of Gcn4 residues 101-134 to Gal11 residues 158-238 is best represented by the entire ensemble as Gcn4 binds in multiple orientations.
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試料調製
詳細
Solution-ID
内容
溶媒系
1
1-1.2 mM [U-15N] Gal11, 2-2.4 mM Gcn4, 20 mM Sodium phosphate, 150 mM Sodium chloride, 1 mM PMSF, 5 mM DTT, 90% H2O/10% D2O
90% H2O/10% D2O
2
2-2.4 mM Gal11, 1-1.2 mM [U-15N] Gcn4, 20 mM Sodium phosphate, 150 mM Sodium chloride, 1 mM PMSF, 5 mM DTT, 90% H2O/10% D2O
90% H2O/10% D2O
3
1-1.2 mM [U-13C; U-15N] Gal11, 2-2.4 mM Gcn4, 20 mM Sodium phosphate, 150 mM Sodium chloride, 1 mM PMSF, 5 mM DTT, 90% H2O/10% D2O
90% H2O/10% D2O
4
1-1.2 mM [U-13C; U-15N] Gal11, 2-2.4 mM Gcn4, 20 mM Sodium phosphate, 150 mM Sodium chloride, 1 mM PMSF, 5 mM DTT, 100% D2O
100% D2O
5
2-2.4 mM Gal11, 1-1.2 mM [U-13C; U-15N] Gcn4, 20 mM Sodium phosphate, 150 mM Sodium chloride, 1 mM PMSF, 5 mM DTT, 90% H2O/10% D2O
90% H2O/10% D2O
試料
濃度 (mg/ml)
単位
構成要素
Isotopic labeling
Conc. range (mg/ml)
Solution-ID
mM
Gal11-1
[U-15N]
1-1.2
1
mM
Gcn4-2
2-2.4
1
20mM
Sodum phosphate-3
1
150mM
Sodum chloride-4
1
1mM
PMSF-5
1
5mM
DTT-6
1
mM
Gal11-7
2-2.4
2
mM
Gcn4-8
[U-15N]
1-1.2
2
20mM
Sodum phosphate-9
2
150mM
Sodum chloride-10
2
1mM
PMSF-11
2
5mM
DTT-12
2
mM
Gal11-13
[U-13C; U-15N]
1-1.2
3
mM
Gcn4-14
2-2.4
3
20mM
Sodum phosphate-15
3
150mM
Sodum chloride-16
3
1mM
PMSF-17
3
5mM
DTT-18
3
mM
Gal11-19
[U-13C; U-15N]
1-1.2
4
mM
Gcn4-20
2-2.4
4
20mM
Sodum phosphate-21
4
150mM
Sodum chloride-22
4
1mM
PMSF-23
4
5mM
DTT-24
4
mM
Gal11-25
2-2.4
5
mM
Gcn4-26
[U-13C; U-15N]
1-1.2
5
20mM
Sodum phosphate-27
5
150mM
Sodum chloride-28
5
1mM
PMSF-29
5
5mM
DTT-30
5
試料状態
イオン強度: 150 / pH: 6.5 / 圧: ambient / 温度: 298 K
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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
900
1
Varian INOVA
Varian
INOVA
800
2
Varian INOVA
Varian
INOVA
600
3
Bruker DMX
Bruker
DMX
500
4
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解析
NMR software
名称
バージョン
開発者
分類
CNS
1.3
Brunger, Adams, Clore, Gros, NilgesandRead
構造決定
NMRPipe
Delaglio, Grzesiek, Vuister, Zhu, PfeiferandBax
データ解析
NMRView
Johnson, OneMoonScientific
データ解析
HADDOCK
2.1
AlexandreBonvin
構造決定
VNMR
Varian
collection
TopSpin
BrukerBiospin
collection
CNS
Brunger, Adams, Clore, Gros, NilgesandRead
精密化
精密化
手法: simulated annealing / ソフトェア番号: 1
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: 13 / 登録したコンフォーマーの数: 13