分子量: 16668.709 Da / 分子数: 1 / 由来タイプ: 組換発現 詳細: Residues 2-124 of the construct correspond to A20:304-426, residues 125-134 to a flexible linker and residues 136-148 to a Biotin Affinity Peptide (BAP),Residues 2-124 of the construct ...詳細: Residues 2-124 of the construct correspond to A20:304-426, residues 125-134 to a flexible linker and residues 136-148 to a Biotin Affinity Peptide (BAP),Residues 2-124 of the construct correspond to A20:304-426, residues 125-134 to a flexible linker and residues 136-148 to a Biotin Affinity Peptide (BAP) 由来: (組換発現) Vaccinia virus (strain Copenhagen) (ウイルス) 株: Copenhagen / 遺伝子: A20R / 発現宿主: Escherichia coli BL21(DE3) (大腸菌) / Variant (発現宿主): -Star / 参照: UniProt: P20995
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実験情報
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実験
実験
手法: 溶液NMR
NMR実験
Conditions-ID
Experiment-ID
Solution-ID
Sample state
Spectrometer-ID
タイプ
1
1
1
isotropic
1
2D 15N-TROSY
1
2
3
isotropic
1
2D 13C-HSQC
1
3
4
isotropic
3
3D HNCO
1
4
4
isotropic
1
3D HN(CA)CB
1
5
3
isotropic
2
3D (H)CCH-TOCSY
1
8
1
isotropic
1
15N-NOESY
1
7
3
isotropic
1
13C-NOESY
1
6
2
isotropic
1
2D-NOESY
1
9
3
isotropic
1
13C-13C-methyl-NOESY
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試料調製
詳細
タイプ
Solution-ID
内容
Label
溶媒系
solution
1
1 mM [U-15N] A20-BAP, 50 mM sodium citrate, 300 mM sodium chloride, 2 mg/L TCEP, 90% H2O/10% D2O
15N-A20-BAP
90% H2O/10% D2O
solution
4
1 mM [U-15N] A20-BAP, 50 mM sodium citrate, 300 mM sodium chloride, 2 mg/L TCEP, 90% H2O/10% D2O
15N13C-A20-BAP-initial
90% H2O/10% D2O
solution
2
1 mM [U-15N] A20-BAP, 50 mM potassium phosphate, 300 mM sodium chloride, 1.2 mM TCEP, 100% D2O
15N-A20-BAP-D2O
100% D2O
solution
3
1 mM [U-15N] A20-BAP, 50 mM potassium phosphate, 300 mM sodium chloride, 1.2 mM TCEP, 90% H2O/10% D2O
13C15N-A20-BAP
90% H2O/10% D2O
試料
濃度 (mg/ml)
構成要素
Isotopic labeling
Solution-ID
1mM
A20-BAP
[U-15N]
1
50mM
sodiumcitrate
naturalabundance
1
300mM
sodiumchloride
naturalabundance
1
2mg/L
TCEP
naturalabundance
1
1mM
A20-BAP
[U-15N]
4
50mM
sodiumcitrate
naturalabundance
4
300mM
sodiumchloride
naturalabundance
4
2mg/L
TCEP
naturalabundance
4
1mM
A20-BAP
[U-15N]
2
50mM
potassiumphosphate
naturalabundance
2
300mM
sodiumchloride
naturalabundance
2
1.2mM
TCEP
naturalabundance
2
1mM
A20-BAP
[U-15N]
3
50mM
potassiumphosphate
naturalabundance
3
300mM
sodiumchloride
naturalabundance
3
1.2mM
TCEP
naturalabundance
3
試料状態
イオン強度: 0.3 M / Label: general / pH: 6 / PH err: 0.1 / 圧: 1 atm / 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
Bruker AVANCE
Bruker
AVANCE
950
1
Bruker AVANCE
Bruker
AVANCE
850
2
Bruker AVANCE
Bruker
AVANCE
600
3
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解析
NMR software
名称
開発者
分類
TopSpin
BrukerBiospin
collection
TopSpin
BrukerBiospin
解析
CcpNmr Analysis
CCPN
chemicalshiftassignment
UNIO
TorstenHerrmann
peakpicking
ARIA
Linge, O'DonoghueandNilges
structurecalculation
精密化
手法: molecular dynamics / ソフトェア番号: 5 / 詳細: water refinement
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: 1000 / 登録したコンフォーマーの数: 20