Text: TRIPLE-RESONANCE NMR SPECTROSCOPY WAS USED TO DETERMINE THE P15 STRUCTURE
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試料調製
詳細
Solution-ID
内容
溶媒系
1
0.2 MM P15, U-15N; 4 MM HEPES, 1 MM DTT, 5 UM EDTA
95% H2O/5% D2O
2
0.2 MM P15, U-15N,13C; 4 MM HEPES, 1 MM DTT, 5 UM EDTA
95% H2O/5% D2O
3
0.2 MM P15, U-15N,13C; 4 MM HEPES, 1 MM DTT, 5 UM EDTA
100% D2O
試料状態
イオン強度: 0.1 / pH: 7.5 / 圧: AMBIENT / 温度: 293 K
結晶化
*PLUS
手法: other / 詳細: NMR
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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 DRX
Bruker
DRX
800
1
Bruker DMX
Bruker
DMX
600
2
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解析
NMR software
名称
バージョン
開発者
分類
XwinNMR
2.1
BRUKER
collection
X-PLOR
3.1
BRUNGER
精密化
MODELLER-4
4
SALI
構造決定
精密化
手法: COMPARATIVE MODELING, RESTRAINT ENERGY MINIMIZATION / ソフトェア番号: 1 詳細: DUE TO THE STRUCTURAL FLEXIBILITY AND INSTABILITY OF P15, ONLY A LIMITED NUMBER OF NMR CONSTRAINTS WERE COLLECTED. BECAUSE P15 SHARES MORE THAN 85% SEQUENCE HOMOLOGY WITH P16, 19 P15 ...詳細: DUE TO THE STRUCTURAL FLEXIBILITY AND INSTABILITY OF P15, ONLY A LIMITED NUMBER OF NMR CONSTRAINTS WERE COLLECTED. BECAUSE P15 SHARES MORE THAN 85% SEQUENCE HOMOLOGY WITH P16, 19 P15 STRUCTURES WERE GENERATED BY USING COMPARATIVE MODELING (MODELLER-4) WITH AN ENSEMBLE OF P16(INK4A) NMR STRUCTURES AS TEMPLATES. THE MODELING STRUCTURES WERE THEN SUBJECTED TO 1400 STEPS OF RESTRAINT ENERGY MINIMIZATION (X-PLOR) WITH THE NMR DATA (672 DISTANCE CONSTRAINTS). THE STRUCTURES THAT SATISFY THE NMR CONSTRAINTS WERE PICKED TO REPRESENT P15(INK4B).
代表構造
選択基準: minimized average structure
NMRアンサンブル
コンフォーマー選択の基準: STRUCTURES WITH THE LEAST RESTRAINT VIOLATIONS, STRUCTURES WITH THE LOWEST ENERGY 計算したコンフォーマーの数: 19 / 登録したコンフォーマーの数: 10