Ensemble Solution structure of the phosphoenolpyruvate-Enzyme I complex from the bacterial hosphotransferase system as determined by NMR residual dipolar couplings and solution X-ray scattering
溶液散乱
NMR実験
Conditions-ID
Experiment-ID
Solution-ID
タイプ
1
1
1
ARTSY
1
2
1
2D 1H-15N HSQC
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試料調製
詳細
内容: 0.4 mM [U-13C; U-15N; U-2H] EIA, 20 mM TRIS, 1 mM EDTA, 100 mM sodium chloride, 2 mM DTT, 90% H2O/10% D2O 溶媒系: 90% H2O/10% D2O
試料
濃度 (mg/ml)
構成要素
Isotopic labeling
Solution-ID
0.4mM
EIA
[U-13C; U-15N; U-2H]
1
20mM
TRIS
naturalabundance
1
1mM
EDTA
naturalabundance
1
100mM
sodiumchloride
naturalabundance
1
2mM
DTT
naturalabundance
1
試料状態
イオン強度: 0.1 / pH: 7.4 / 圧: ambient / 温度: 310 K
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データ収集
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: 800 MHz
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解析
NMR software
名称
開発者
分類
Sparky
Goddard
chemicalshiftassignment
Xplor-NIH
Schwieters, Kuszewski, TjandraandClore
精密化
NMRPipe
Delaglio, Grzesiek, Vuister, Zhu, PfeiferandBax
解析
TopSpin
BrukerBiospin
collection
精密化
手法: simulated annealing / ソフトェア番号: 2 詳細: The deposited structures represent two-membered ensembles with varying weights which must be considered together to properly reproduce the RDC and SAXS data. The first members of 10 ensembles ...詳細: The deposited structures represent two-membered ensembles with varying weights which must be considered together to properly reproduce the RDC and SAXS data. The first members of 10 ensembles are reported in models 1-10 which were allowed rigid-body motion of subunits, arbitrary motion of linker regions and torsion degrees of freedom for sidechains. The second member of each ensemble is represented in model 11, which was fixed in the calculations. The ten pairs of weights for the ten ensemble members are Ensemble 1: MODEL 1, weight: 0.503 MODEL 11, weight: 0.497 Ensemble 2: MODEL 2, weight: 0.518 MODEL 11, weight: 0.482 Ensemble 3: MODEL 3, weight: 0.526 MODEL 11, weight: 0.474 Ensemble 4: MODEL 4, weight: 0.520 MODEL 11, weight: 0.480 Ensemble 5: MODEL 5, weight: 0.508 MODEL 11, weight: 0.492 Ensemble 6: MODEL 6, weight: 0.531 MODEL 11, weight: 0.469 Ensemble 7: MODEL 7, weight: 0.498 MODEL 11, weight: 0.501 Ensemble 8: MODEL 8, weight: 0.524 MODEL 11, weight: 0.476 Ensemble 9: MODEL 9, weight: 0.514 MODEL 11, weight: 0.486 Ensemble 10: MODEL 10, weight: 0.526 MODEL 11, weight: 0.474
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: 200 / 登録したコンフォーマーの数: 11 / Maximum torsion angle constraint violation: 19.4 °