Mass: 8966.376 Da / Num. of mol.: 1 / Fragment: C-TERMINAL DOMAIN Source method: isolated from a genetically manipulated source Source: (gene. exp.) Aquifex aeolicus (bacteria) / Gene: rpoN / Plasmid: pet21a / Production host: Escherichia coli (E. coli) / Strain (production host): Rosetta (BL21 (DE3) with pLysS) / References: UniProt: O66858
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Experimental details
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Experiment
Experiment
Method: SOLUTION NMR
NMR experiment
Conditions-ID
Experiment-ID
Solution-ID
Type
1
1
1
3D 15N-separated NOESY
1
2
1
3D N15-seperated TOCSY
1
3
2
HNCA-J
1
4
2
(H)CCH-TOCSY
1
5
2
3D 13C-separated NOESY
1
6
4
2D C13-HSQC
NMR details
Text: Other experiments performed and used for chemical shift assignments and obtaining restraints: CBCA(CO)NH, DQF-COSY, CC(CO)NH, H/D Exchange via N15-HSQC, HNHA - J couplings
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Sample preparation
Details
Solution-ID
Contents
Solvent system
1
0.8 mM Sigma-54 U-15N; 50 mM Hepes; 250 mM NaCl; 1 mM EDTA; 90% H20; 10% D20
90% H20; 10% D20
2
0.8 mM Sigma-54 U-15N U-13C; 50 mM Hepes; 250 mM NaCl; 1 mM EDTA; 90% H20; 10% D20
90% H20; 10% D20
3
0.8 mM Sigma-54 U-15; 50 mM Hepes; 250 mM NaCl; 1 mM EDTA; 5% H20; 95% D20
5% H20; 95% D20
4
0.8 mM Sigma-54 U-15N U-10% 13C; 50 mM Hepes; 250 mM NaCl; 1 mM EDTA; 90% H20; 10% D20
90% H20; 10% D20
Sample conditions
Ionic strength: 50 mM Hepes; 250 mM NaCl / pH: 6.8 / Pressure: ambient / Temperature: 298 K
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NMR measurement
Radiation
Protocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M
Radiation wavelength
Relative weight: 1
NMR spectrometer
Type: Bruker DRX / Manufacturer: Bruker / Model: DRX / Field strength: 600 MHz
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Processing
NMR software
Name
Version
Developer
Classification
NMRPipe
2.3
Delaglioetal. (1995)
processing
NMRView
5.0.4
Johnson & Blevins (1994)
dataanalysis
DYANA
1.5
Guntertetal. (1997)
structuresolution
DYANA
1.5
Guntertetal. (1997)
refinement
Refinement
Method: torsion angle dynamics / Software ordinal: 1 Details: Structures are based on a total 1064 restraints: 956 distance and 108 dihedral angle
NMR representative
Selection criteria: lowest dyana target function
NMR ensemble
Conformer selection criteria: target function / Conformers calculated total number: 150 / Conformers submitted total number: 20
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