分子量: 17976.117 Da / 分子数: 1 / 由来タイプ: 組換発現 由来: (組換発現) Mycobacterium tuberculosis (結核菌) 解説: original pET16bTev vector was double NcoI/BlpId digested and cloned into the modified pKM263 (6xHis tag ProtGB1-TEV between Nde1 and Xho1) vector 遺伝子: ptpA, Rv2234, MT2293, MTCY427.15 / プラスミド: pKM263 / 発現宿主: Escherichia coli (大腸菌) / 株 (発現宿主): BL21(DE3) / Variant (発現宿主): pLysS 参照: UniProt: P65716, UniProt: P9WIA1*PLUS, protein-tyrosine-phosphatase
-
実験情報
-
実験
実験
手法: 溶液NMR / 詳細: SORTED ON LOWEST ENERGY AFTER ARIA WATER REFINEMENT
NMR実験
Conditions-ID
Experiment-ID
Solution-ID
タイプ
1
1
2
2D 1H-15N HSQC
1
2
1
2D 1H-13C HSQC
1
3
1
3D HN(CA)CB
1
4
1
3D (H)CCH-TOCSY
1
5
1
3D (H)C(NC)H
1
6
1
3D HCD(CG)CB-TOCSY
1
7
1
3D (H)CC(CO)NH-TOCSY
1
8
1
3D (H)CB(CG)CCH-TOCSY
1
9
1
3D 1H-13C NOESY aliphatic
1
10
1
3D 1H-13C NOESY aromatic
1
11
2
3D 1H-15N NOESY
1
12
2
2D (T1,T2,HetNOE) 1H-15N HSQC
1
13
2
2D (IPAP) 1H-15N HSQC
1
14
2
2D (IPAP) 1H-15N HSQC
1
15
2
3D HNHA
NMR実験の詳細
Text: SORTED ON LOWEST ENERGY AFTER ARIA WATER REFINEMENT
-
試料調製
詳細
Solution-ID
内容
溶媒系
1
1.2 mM [U-100% 13C; U-100% 15N] mptpa, 50 mM arginine / glutamine, 25 mM HEPES, 10 mM DTT, 90% H2O/10% D2O
90% H2O/10% D2O
2
1.0 mM [U-100% 15N] mptpa, 50 mM arginine / glutamine, 25 mM HEPES, 10 mM DTT, 90% H2O/10% D2O
90% H2O/10% D2O
試料
濃度 (mg/ml)
構成要素
Isotopic labeling
Solution-ID
1.2mM
mptpa-1
[U-100% 13C; U-100% 15N]
1
50mM
arginine / glutamine-2
1
25mM
HEPES-3
1
10mM
DTT-4
1
1.0mM
mptpa-5
[U-100% 15N]
2
50mM
arginine / glutamine-6
2
25mM
HEPES-7
2
10mM
DTT-8
2
試料状態
イオン強度: 0 / pH: 7.0 / 圧: ambient / 温度: 303 K
-
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
900
2
Bruker Avance
Bruker
AVANCE
800
3
Bruker Avance
Bruker
AVANCE
600
4
-
解析
NMR software
名称
バージョン
開発者
分類
TopSpin
2.1
BrukerBiospin
collection
TopSpin
2.1
BrukerBiospin
解析
Sparky
3.114
Goddard
chemicalshiftassignment
Sparky
3.114
Goddard
データ解析
Sparky
3.114
Goddard
peakpicking
CARA
1.8.4.2
KellerandWuthrich
chemicalshiftassignment
CARA
1.8.4.2
KellerandWuthrich
データ解析
CARA
1.8.4.2
KellerandWuthrich
peakpicking
CYANA
3.9
Guntert, MumenthalerandWuthrich
構造決定
CNS
1.1
Brunger, Adams, Clore, Gros, NilgesandRead
構造決定
ARIA
1.2
Linge, O'DonoghueandNilges
構造決定
CYANA
3.9
Guntert, MumenthalerandWuthrich
精密化
CNS
1.1
Brunger, Adams, Clore, Gros, NilgesandRead
精密化
ARIA
1.2
Linge, O'DonoghueandNilges
精密化
精密化
手法: SIMULATED ANNEALING, TORSION ANGLE DYNAMICS, MOLECULAR DYNAMICS ソフトェア番号: 1 詳細: SIMULATED ANNEALING WITH TORSION ANGLE DYNAMICS, STRUCTURE DETERMINATION WITH AUTOMATED NOE ASSIGNMENT USING CYANA, FINAL STRUCTURE REFINEMENT IN EXPLICIT WATER WITH ADDITIONAL DIFFUSION ...詳細: SIMULATED ANNEALING WITH TORSION ANGLE DYNAMICS, STRUCTURE DETERMINATION WITH AUTOMATED NOE ASSIGNMENT USING CYANA, FINAL STRUCTURE REFINEMENT IN EXPLICIT WATER WITH ADDITIONAL DIFFUSION ANISOTROPY DATA USING ARIA PROTOCOLS WITH CNS.
NMR constraints
NOE constraints total: 4821 / NOE intraresidue total count: 815 / NOE long range total count: 1611 / NOE medium range total count: 1103 / NOE sequential total count: 1292 / Hydrogen bond constraints total count: 138
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: target function / 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: 20