Mass: 10025.209 Da / Num. of mol.: 1 / Fragment: UNP residues 11-94 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Drosophila melanogaster (fruit fly) / Gene: MRG15, CG6363 / Production host: Escherichia coli (E. coli) / References: UniProt: Q9Y0I1
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Experimental details
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Experiment
Experiment
Method: SOLUTION NMR Details: 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 ...Details: 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 experiment
Conditions-ID
Experiment-ID
Solution-ID
Type
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
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Sample preparation
Details
Solution-ID
Contents
Solvent system
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
Conformer selection criteria: structures with the least restraint violations Conformers calculated total number: 50 / Conformers submitted total number: 10
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