手法: 溶液NMR 詳細: The data from this entry were used to determine structure of Blo t 19. Due to Proline isomerization, Blo t 19 spectrum has degenerate resonances for a stretch of six residues in its C- ...詳細: The data from this entry were used to determine structure of Blo t 19. Due to Proline isomerization, Blo t 19 spectrum has degenerate resonances for a stretch of six residues in its C-terminus. These two populations have varying intensities and are unambiguously assigned as major and minor population. Solution structure is calculated for the Major population.
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 HNCO
1
4
1
3DHN(CA)CO
1
5
1
3D HNCA
1
6
1
3D HN(CA)CB
1
7
1
3DCBCA(CO)NH
1
8
1
3D HNHA
1
9
1
3D (H)CCH-COSY
1
10
1
3D (H)CCH-TOCSY
1
11
1
2D-(HB)CB(CGCD)HD
1
12
1
2D-(HB)CB(CGCDCE)HE
1
13
3
2D-TOCSY
1
14
2
3D 1H-15N TOCSY
1
15
3
2D 1H-1H NOESY
1
16
1
3D 1H-13C NOESYHSQC
1
17
1
3D 1H-15N NOESYHSQC
1
18
4
2D 1H-15N IPAPHSQC
NMR実験の詳細
Text: NMR data was acquired at 295K using Shigemi NMR tubes.
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試料調製
詳細
Solution-ID
内容
溶媒系
1
1 mM [U-100% 13C; U-100% 15N] Blo t 19-1, 50 mM potassium phosphate-2, 100 mM sodium chloride-3, 90% H2O/10% D2O
90% H2O/10% D2O
2
1 mM [U-100% 15N] Blo t 19-4, 50 mM potassium phosphate-5, 100 mM sodium chloride-6, 90% H2O/10% D2O
90% H2O/10% D2O
3
1 mM Blo t 19-7, 50 mM potassium phosphate-8, 100 mM sodium chloride-9, 90% H2O/10% D2O
90% H2O/10% D2O
4
1 mM [U-100% 15N] Blo t 19-10, 50 mM potassium phosphate-11, 100 mM sodium chloride-12, 15 mg/ml Pf1 phage-13, 90% H2O/10% D2O
90% H2O/10% D2O
試料
濃度 (mg/ml)
構成要素
Isotopic labeling
Solution-ID
1mM
Blo t 19-1
[U-100% 13C; U-100% 15N]
1
50mM
potassium phosphate-2
1
100mM
sodium chloride-3
1
1mM
Blo t 19-4
[U-100% 15N]
2
50mM
potassium phosphate-5
2
100mM
sodium chloride-6
2
1mM
Blo t 19-7
3
50mM
potassium phosphate-8
3
100mM
sodium chloride-9
3
1mM
Blo t 19-10
[U-100% 15N]
4
50mM
potassium phosphate-11
4
100mM
sodium chloride-12
4
15mg/mL
Pf1 phage-13
4
試料状態
pH: 6.0 / 圧: ambient / 温度: 295 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 Avance
Bruker
AVANCE
500
1
Bruker Avance
Bruker
AVANCE
600
2
Bruker Avance
Bruker
AVANCE
800
3
Bruker Avance
Bruker
AVANCE
850
4
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解析
NMR software
名称
バージョン
開発者
分類
TopSpin
3
BrukerBiospin
collection
TopSpin
3
BrukerBiospin
解析
Sparky
Goddard
chemicalshiftassignment
Sparky
Goddard
peakpicking
CYANA
3.9
Guntert, MumenthalerandWuthrich
構造決定
CYANA
3.9
Guntert, MumenthalerandWuthrich
geometryoptimization
PSVS
1.5
BhattacharyaandMontelione
構造検証
X-PLOR NIH
2.34
Schwieters, Kuszewski, TjandraandClore
精密化
CYANA
Guntert, MumenthalerandWuthrich
精密化
精密化
手法: torsion angle dynamics, DGSA-distance geometry simulated annealing ソフトェア番号: 1 詳細: Initial structure ensemble was calculated by semi-automated NOESY assignment by CYANA. The assignments were manually verified in Sparky and final structure annealing was performed in CYANA. ...詳細: Initial structure ensemble was calculated by semi-automated NOESY assignment by CYANA. The assignments were manually verified in Sparky and final structure annealing was performed in CYANA. Structure and restraints from CYANA were imported in Xplor-NIH for explicit water refinement.
NMR constraints
NOE constraints total: 1140 / NOE intraresidue total count: 212 / NOE long range total count: 294 / NOE medium range total count: 314 / NOE sequential total count: 320 / Disulfide bond constraints total count: 12 / Hydrogen bond constraints total count: 54 / Protein chi angle constraints total count: 0 / Protein other angle constraints total count: 43 / Protein phi angle constraints total count: 39 / Protein psi angle constraints total count: 39
代表構造
選択基準: lowest energy
NMRアンサンブル
Average torsion angle constraint violation: 0.08688 ° 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: 400 / 登録したコンフォーマーの数: 20 / Maximum torsion angle constraint violation: 4.9 ° / Maximum upper distance constraint violation: 0.7 Å / 代表コンフォーマー: 1 / Torsion angle constraint violation method: PSVS
NMR ensemble rms
Distance rms dev: 0.02648 Å / Distance rms dev error: 0.02581 Å