1.06 mM [U-100% 13C; U-100% 15N] GmR137, THiS protein, 90% H2O/10% D2O
90% H2O/10% D2O
2
1.26 mM [U-10% 13C; U-99% 15N] GmR137, THiS protein, 90% H2O/10% D2O
90% H2O/10% D2O
試料
濃度 (mg/ml)
構成要素
Isotopic labeling
Solution-ID
1.06mM
GmR137, THiSprotein
[U-100% 13C; U-100% 15N]
1
1.26mM
GmR137, THiSprotein
[U-10% 13C; U-99% 15N]
2
試料状態
Conditions-ID
イオン強度
pH
圧 (kPa)
温度 (K)
1
5mMCaCl2, 100mMNaCl
6.5
ambient
293K
2
5mMCaCl2, 100mMNaCl
6.5
ambient
293K
-
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
800
1
Varian INOVA
Varian
INOVA
600
2
Varian INOVA
Varian
INOVA
500
3
-
解析
NMR software
名称
バージョン
開発者
分類
CNS
2.0.6
Brunger, Adams, Clore, Gros, NilgesandRead
精密化
AutoStructure
2.2.1
Huang, Tejero, PowersandMontelione
構造決定
CYANA
2.1
Guntert, MumenthalerandWuthrich
構造決定
AutoAssign
2.4.0
Zimmerman, Moseley, KulikowskiandMontelione
chemicalshiftassignment
精密化
手法: simulated annealing / ソフトェア番号: 1 詳細: The structure was obtained using triple resonance NMR spectroscopy. GFT-NMR experiments were used for backbone resonance assignments and conventional 3D TOCSY and COSY experiments were used ...詳細: The structure was obtained using triple resonance NMR spectroscopy. GFT-NMR experiments were used for backbone resonance assignments and conventional 3D TOCSY and COSY experiments were used to obtain sidechain resonance assignments. Automated NOESY assignments were made using AUTOSTRUCTURE and CYANA-2.1. Dihedral angle constraints were obtained from TALOS. The structure calculation was done excluding 5 HIS from 8-residue C-terminal tag (LEHHHHHH). Completeness of assignments excluding 5-HIS: Backbone - 100%, Sidechain (aliphatic) - 99% Sidechain (aromatic) - 96%. The assignments were validated using AVS software. Final structure quality factros (excluding 5 HIS) determined using PSVS-v1.3: Ordered residues are defiend as: 1-13, 18-29, 32-63. (a) RMSD (ordered residues) all backbone atoms: 0.7A and heavy atoms: 1.2A. (b) Ramachandran statistics for ordered residues: Most favored region: 82.8%, additionally allowed regions: 17.0% generously allowed region: 0.1%, disallowed regions: 0.1%. (c) Procheck scores for ordered residues (RAW/Z-): Phi/psi -0.58/-1.97, all -0.48/-2.84, (d) MolProbity clashscores (RAW/Z-) 17.03/-1.40, (e) RPF scores for the goodness of the fit to NOESY data: Recall - 0.99, Precision - 0.942, F-measure - 0.966 and final dp score - 0.88. (f) Number of close contacts for 20 models: 6, RMS deviation for bond angles - 0.6deg, RMS deviation for bond lengths - 0.009A.
NMR constraints
NOE constraints total: 969 / NOE intraresidue total count: 144 / NOE long range total count: 269 / NOE medium range total count: 207 / NOE sequential total count: 349 / 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: 140 / 登録したコンフォーマーの数: 20 / Maximum upper distance constraint violation: 0.2 Å / Torsion angle constraint violation method: PSVS software