手法: 溶液NMR 詳細: The oligomeric states of the Fr822A were verified by correlation time measurements and concentration dependent studies. The results indicated that the Fr822A self-associates weakly at protein ...詳細: The oligomeric states of the Fr822A were verified by correlation time measurements and concentration dependent studies. The results indicated that the Fr822A self-associates weakly at protein concentration below 1 mM. Using the residual dipolar couplings as well as chemical shift perturbations on dilution, we constructed dimeric models for Fr822A. Possible inter-subunit NOEs were identified based on these models. In cases where corresponding NOEs were ambiguous with intra-molecular NOEs, they were eliminated as constraints and the protein structure recalculated. The absence of structural variation and the failure to observed other predicted inter-subunit NOEs validated use of all NOEs as intra-subunit constraints.
NMR実験
Conditions-ID
Experiment-ID
Solution-ID
タイプ
1
1
1
2D 1H-15N HSQC
1
2
1
2D 1H-13C HSQC aliphatic
1
3
1
2D 1H-13C HSQC aromatic
1
4
1
3DCBCA(CO)NH
1
5
1
3DC(CO)NH
1
6
1
3D HNCO
1
7
1
3D HNCA
1
8
1
3D HN(CA)CB
1
9
1
3DHBHA(CO)NH
1
10
1
3D (H)CCH-TOCSY
1
11
1
3D 1H-15N NOESY
1
12
1
3D 1H-13C NOESY aliphatic
1
13
1
3D 1H-13C NOESY aromatic
1
14
1
2D 1H-15N NH-J-Modulation-HSQC
1
15
2
2D 1H-15N NH-J-Modulation-HSQC
1
16
3
2D 1H-15N NH-J-Modulation-HSQC
-
試料調製
詳細
Solution-ID
内容
溶媒系
1
0.9 mM [U-100% 13C; U-100% 15N] Fr822A, 20 mM sodium phosphate, 200 mM sodium chloride, 5 mM DTT, 95% H2O/5% D2O
95% H2O/5% D2O
2
0.9 mM [U-100% 15N] Fr822A, 20 mM sodium phosphate, 200 mM sodium chloride, 12 mg Pf1 phage, 5 mM DTT, 95% H2O/5% D2O
95% H2O/5% D2O
3
0.9 mM [U-100% 15N] Fr822A, 20 mM sodium phosphate, 200 mM sodium chloride, 4.2 % C12E5, 5 mM DTT, 95% H2O/5% D2O