根拠: The protein's NMR rotational correlation time increased upon addition of nanodiscs indicating interactions between the molecules. Furthermore, ultra-centrifugation, fluorescence, and ...根拠: The protein's NMR rotational correlation time increased upon addition of nanodiscs indicating interactions between the molecules. Furthermore, ultra-centrifugation, fluorescence, and mutagenesis were used to demonstrate the association with vesicles in the orientation determined by NMR.
タイプ: solution 内容: 20 mM Tris-HCl, 300 mM NaCl, 0.02 % sodium azide, 93 % H2O, 7 % [U-100% 2H] D2O, 90 uM 2H, 13C, 15N MT1-MMP hemopexin-like domain, 180 uM MSP1D1, 14.4 mM PX4, 93% H2O/7% D2O 詳細: 300 mM NaCl was used to reduce affinity of HPX to the nanodisc and recover sample. Shaped NMR tubes were used to obtain better spectra. Label: 2H, 13C, 15N / 溶媒系: 93% H2O/7% D2O
試料
濃度 (mg/ml)
構成要素
Isotopic labeling
Solution-ID
20mM
Tris-HCl
naturalabundance
1
300mM
NaCl
naturalabundance
1
0.02 %
sodiumazide
naturalabundance
1
93 %
H2O
naturalabundance
1
7 %
D2O
[U-100% 2H]
1
90uM
MT1-MMP hemopexin-like domain
2H, 13C, 15N
1
180uM
MSP1D1
naturalabundance
1
14.4mM
PX4
naturalabundance
1
試料状態
詳細: MT1-MMP hemopxin-like domain was along with MSP1D1 and PX4 lipids were in a buffer containing 300 mM NaCl, 0.02% sodium azide, 93% water, 7% D2O, 20 mM Tris-HCl pH 7.2 イオン強度: 300 mM / Label: NMR Buffer / pH: 7.2 / 圧: 1 atm / 温度: 303 K
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NMR測定
NMRスペクトロメーター
タイプ: Bruker AVANCE III / 製造業者: Bruker / モデル: AVANCE III / 磁場強度: 800 MHz
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解析
NMR software
名称
バージョン
開発者
分類
TopSpin
3.2
Bruker
解析
Analysis
2.4
CCPN
データ解析
HADDOCK
HADDOCK2.1
Bonvin
structurecalculation
NAMD
NAMD2.1withCUDAGPUprocessing
Theoretical and Computational Biophysics Group in the Beckman Institute for Advanced Science and Technology at the University of Illinois at Urbana-Champaign
structurecalculation
NAMD
NAMD2.1withCUDAGPUprocessing
Theoretical and Computational Biophysics Group in the Beckman Institute for Advanced Science and Technology at the University of Illinois at Urbana-Champaign