MODEL 1 IS PHOSPHORYLATED-HIS IN CHAIN B OF IIAMAN. MODEL 2 IS A PHOSPHORYL TRANSITION STATE ...MODEL 1 IS PHOSPHORYLATED-HIS IN CHAIN B OF IIAMAN. MODEL 2 IS A PHOSPHORYL TRANSITION STATE INVOLVING A PENTACOORDINATE PHOSPHORYL GROUP IN A TRIGONAL BIPYRAMIDAL GEOMETRY WITH THE DONOR (NE2 OF HIS10 OF CHAIN B OF IIAMAN) AND ACCEPTOR (ND1 OF HIS175 OF IIBMAN, CHAIN C) IN APICAL POSITIONS. THE STRUCTURE OF THE TRANSITION STATE IS DERIVED FROM MODEL 1. IN THIS TRANSITION STATE WITH PARTIAL DISSOCIATIVE CHARACTER THE N-P DISTANCE IS 2.5 A. THE COORDINATES OF MODEL 1 ARE IDENTICAL TO THOSE OF 2JZN (IIAMANH10E-IIBMAN COMPLEX EXCEPT THAT GLU10 OF CHAIN B OF IIAMANNOSE HAS BEEN REPLACED BY PHOSPHORYLATED HISTIDINE. MODEL 2 IS DERIVED FROM MODEL 1 BY RESTRAINED MINIMIZATION IN WHICH MINIMAL SHIFTS IN THE BACKBONE COORDINATES (AND ASSOCIATED SIDE CHAINS) OF RESIDUES 8-12 OF CHAIN B OF IIAMANNOSE AND RESIDUES 173-177 OF IIBMANNOSE (CHAIN C) ARE ALLOWED TO OCCUR IN ORDER TO ACCOMMODATE THE STEREOCHEMICAL REQUIREMENTS OF THE TRANSITION STATE. SEE PUBLICATION FOR DETAILS.
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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
タイプ
1
1
1
15N-edited/13C-edited NOE
1
2
2
12C-filtered/13C-separated NOE
NMR実験の詳細
Text: Triple resonance experiments for assignment. Quantitative J correlation experiments for heteronuclear scalar couplings. 3D and 2D isotope filtered/isotope-separated NOE experiments for intermolecular NOEs.
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試料調製
詳細
Solution-ID
内容
溶媒系
1
0.5-1 mM IIAMan(H10E)+IIBMan, 90% H2O/10% D2O
90% H2O/10% D2O
2
0.5-1 mM IIAMan(H10E)+IIBMan, 100% D2O
100% D2O
試料
濃度 (mg/ml)
構成要素
Isotopic labeling
Solution-ID
0.5mM
IIAMan(H10E)+IIBMan
1
20mM
sodiumphosphate
naturalabundance
1
0.01 %
sodiumazide
naturalabundance
1
0.5mM
IIAMan(H10E)+IIBMan
2
20mM
sodiumphosphate
naturalabundance
2
0.01 %
sodiumazide
naturalabundance
2
試料状態
イオン強度: 20 mM phosphate / pH: 6.5 / 圧: ambient / 温度: 303 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 DMX500
Bruker
DMX500
500
1
Bruker DMX600
Bruker
DMX600
600
2
Bruker DRX600
Bruker
DRX600
600
3
Bruker DRX800
Bruker
DRX800
800
4
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解析
NMR software
名称
バージョン
開発者
分類
X-PLOR NIH
2.18.1
Schwieters, Kuszewski, andClore
構造決定
X-PLOR NIH
2.18.1
Schwieters, Kuszewski, andClore
精密化
精密化
手法: conjoined rigid body, torsion angle simulated annealing ソフトェア番号: 2 詳細: COORDINATES OF IIAMAN (1PDO) AND IIBMAN (2JZH) ARE TREATED AS RIGID BODIES WITH THE INTERFACIAL SIDE CHAINS GIVEN TORSIONAL DEGREES OF FREEDOM. RESIDUES 130- 134 OF IIAMAN ARE ALSO GIVEN ...詳細: COORDINATES OF IIAMAN (1PDO) AND IIBMAN (2JZH) ARE TREATED AS RIGID BODIES WITH THE INTERFACIAL SIDE CHAINS GIVEN TORSIONAL DEGREES OF FREEDOM. RESIDUES 130- 134 OF IIAMAN ARE ALSO GIVEN TORSIONAL DEGREES OF FREEDOM SINCE INTERMOLECULAR NOES WERE OBSERVED INVOLVED THESE RESIDUES ALTHOUGH THEY ARE NOT VISIBLE IN THE ELECTRON DENSITY MAP OF THE X-RAY STRUCTURE OF FREE IIAMAN. FURTHER DETAILS IN PUBLICATION. THE COORDINATES of 1VSQ ARE DERIVED DIRECTLY FROM THOSE OF 2JZN, THE IIAMAN(H10E)-IIBMAN COMPLEX.
NMRアンサンブル
コンフォーマー選択の基準: restrained regularized mean 計算したコンフォーマーの数: 120 / 登録したコンフォーマーの数: 2