Text: All 2D and 3D experiments were TROSY based. For the RDC data collection, the TROSY/Semi-TROSY method was used.
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試料調製
詳細
内容: 10 % lyso myristoyl phosphatidylglycerol, 10 % [U-2H] D2O, 100 mM imidazole, 250 uM [U-100% 15N] APP_C99, 1 mM EDTA, 90% H2O/10% D2O 溶媒系: 90% H2O/10% D2O
試料
濃度 (mg/ml)
構成要素
Isotopic labeling
Solution-ID
10 %
lyso myristoyl phosphatidylglycerol-1
1
10 %
D2O-2
[U-2H]
1
100mM
imidazole-3
1
250uM
APP_C99-4
[U-100% 15N]
1
1mM
EDTA-5
1
10 %
lyso myristoyl phosphatidylglycerol-6
2
10 %
D2O-7
[U-2H]
2
100mM
imidazole-8
2
250uM
APP_C99-9
[U-100% 15N]
2
1mM
EDTA-10
2
10 %
lyso myristoyl phosphatidylglycerol-11
3
10 %
D2O-12
[U-2H]
3
100mM
imidazole-13
3
250uM
APP_C99-14
[U-100% 15N]
3
1mM
EDTA-15
3
10 %
lyso myristoyl phosphatidylglycerol-16
4
10 %
D2O-17
[U-2H]
4
100mM
imidazole-18
4
250uM
APP_C99-19
[U-100% 15N]
4
1mM
EDTA-20
4
10 %
lyso myristoyl phosphatidylglycerol-21
5
10 %
D2O-22
[U-2H]
5
100mM
imidazole-23
5
250uM
APP_C99-24
[U-15N; U-2H]
5
7 %
Acrylamide-25
5
0.5 %
AMPS-26
5
1mM
EDTA-27
5
9 %
lyso myristoyl phosphatidylglycerol-28
6
1mM
APP_C99-29
[U-100% 13C; U-100% 15N; U-80% 2H]
6
10 %
D2O-30
[U-2H]
6
2.5mM
EDTA-31
6
250mM
imidazole-32
6
試料状態
pH: 6.5 / 圧: ambient / 温度: 318 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 AMX
Bruker
AMX
800
1
Bruker AMX
Bruker
AMX
600
2
Varian INOVA
Varian
INOVA
900
3
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解析
NMR software
名称
バージョン
開発者
分類
X-PLOR NIH
2.24
Schwieters, Kuszewski, TjandraandClore
構造決定
X-PLOR NIH
2.24
Schwieters, Kuszewski, TjandraandClore
精密化
NMRDraw
Linux9
Delaglio, Grzesiek, Vuister, Zhu, PfeiferandBax
データ解析
Sparky
3.114
Goddard
データ解析
Sparky
3.114
Goddard
chemicalshiftassignment
Sparky
3.114
Goddard
peakpicking
NMRPipe
Linux9
Delaglio, Grzesiek, Vuister, Zhu, PfeiferandBax
解析
TALOS
Linux9
Cornilescu, DelaglioandBax
geometryoptimization
TALOS
Linux9
Cornilescu, DelaglioandBax
構造決定
ProcheckNMR
3.5.4
LaskowskiandMacArthur
データ解析
ProcheckNMR
3.5.4
LaskowskiandMacArthur
構造決定
TopSpin
3
BrukerBiospin
collection
精密化
手法: molecular dynamics, simulated annealing / ソフトェア番号: 1 詳細: RESTRAINED MOLECULAR DYNAMICS WITH SIMULATED ANNEALING, FOLLOWED BY POWELL ENERGY MINIMIZATION
NMR constraints
NOE constraints total: 65 / NOE intraresidue total count: 0 / NOE long range total count: 0 / NOE medium range total count: 22 / NOE sequential total count: 43 / Hydrogen bond constraints total count: 13 / Protein chi angle constraints total count: 0 / Protein other angle constraints total count: 0 / Protein phi angle constraints total count: 33 / Protein psi angle constraints total count: 32
代表構造
選択基準: closest to the average
NMRアンサンブル
Average torsion angle constraint violation: 0.351 ° 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: 100 / 登録したコンフォーマーの数: 30 / Maximum lower distance constraint violation: 0 Å / Maximum torsion angle constraint violation: 0.56 ° / Maximum upper distance constraint violation: 0 Å / Torsion angle constraint violation method: NIH Xplor
NMR ensemble rms
Distance rms dev: 0 Å / Distance rms dev error: 0 Å