Mass: 24483.572 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Escherichia coli (E. coli) / Description: sample is uniformly 15N labeled / Production host: Escherichia coli (E. coli) / Strain (production host): BL21(DE3) / References: GenBank: 1845291569
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Experimental details
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Experiment
Experiment
Method
SOLUTION NMR
SOLUTION SCATTERING
NMR experiment
Conditions-ID
Experiment-ID
Solution-ID
Type
1
1
1
2D 1H-15N TROSY-HSQC
1
2
1
2D 1H-15N HSQC
1
3
1
2D 1H-15N TROSY-HSQC
1
4
1
2D 1H-15N HSQC
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Sample preparation
Details
Solution-ID
Contents
Solvent system
1
0.5 mM [U-100% 15N] RNA (76-MER), 10 mM sodium phosphate, 80 mM sodium chloride, 5 mM magnesium chloride, 0.1 mM EDTA, 90% H2O/10% D2O
90% H2O/10% D2O
2
0.41 mM [U-100% 15N] RNA (76-MER), 10 mM sodium phosphate, 150 mM sodium chloride, 5 mM magnesium chloride, 0.1 mM EDTA, 100% H2O
100% H2O
Sample
Conc. (mg/ml)
Component
Isotopic labeling
Solution-ID
0.5mM
RNA (76-MER)
[U-100% 15N]
1
10mM
sodiumphosphate
1
80mM
sodiumchloride
1
5mM
magnesiumchloride
1
0.1mM
EDTA
1
0.41mM
RNA (76-MER)
[U-100% 15N]
2
10mM
sodiumphosphate
2
150mM
sodiumchloride
2
5mM
magnesiumchloride
2
0.1mM
EDTA
2
Sample conditions
Conditions-ID
Ionic strength
pH
Pressure (kPa)
Temperature (K)
1
0.155
6.8
ambientatm
298K
2
0.225
7.0
ambientatm
298K
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Data collection
NMR spectrometer
Type
Manufacturer
Model
Field strength (MHz)
Spectrometer-ID
Bruker DRX
Bruker
DRX
800
1
Bruker DMX
Bruker
DMX
500
2
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Processing
NMR software
Name
Version
Developer
Classification
X-PLOR NIH
2.9.14
Schwieters, Kuszewski, TjandraandClore
refinement
CNS
1
Brunger, Adams, Clore, Gros, NilgesandRead
refinement
Refinement
Method: simulated annealing, torsion angle dynamics / Software ordinal: 1 Details: tRNAVal solution structure is obtained by refinement of the tRNAPhe based homology model against RDC data (base N-H RDCs) from Pf1 and self-alignment (MSA), and SAXS data. Totals of 24 Pf1 ...Details: tRNAVal solution structure is obtained by refinement of the tRNAPhe based homology model against RDC data (base N-H RDCs) from Pf1 and self-alignment (MSA), and SAXS data. Totals of 24 Pf1 RDCs and 20 MSA RDcs were fitted. During the refinement, the local geometry was kept close to the tRNAPhe based homology model using a set of non-crystallographic symmetry restraint terms. The NCS potentials included: 70 sequential i/i+1 terms and 34 additional terms for base pairs and triplets. The experimental SAXS data between q=0.03 and 0.35 A^-1 were fitted as well.
NMR representative
Selection criteria: lowest energy
NMR ensemble
Conformer selection criteria: structures with the lowest energy Conformers calculated total number: 3 / Conformers submitted total number: 1
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