Mass: 7095.323 Da / Num. of mol.: 1 / Source method: obtained synthetically / Details: In vitro transcription by T7 RNA polymerase
#2: Protein
NUCLEOLINRBD12 / PROTEIN C23
Mass: 19365.744 Da / Num. of mol.: 1 / Fragment: TWO N-TERMINAL RBD DOMAINS Source method: isolated from a genetically manipulated source Source: (gene. exp.) Mesocricetus auratus (golden hamster) / Plasmid: PET15-B / Production host: Escherichia coli (E. coli) / References: UniProt: P08199
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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 13C-separated NOESY
1
2
1
13C-12C filtered 3D
1
3
2
3D 15N-separated NOESY
1
4
3
2D NOESY
2
5
3
2D NOESY
1
6
4
3D 13C-separated NOESY
NMR details
Text: The structure was determined using triple-resonance NMR spectroscopy
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Sample preparation
Details
Solution-ID
Contents
Solvent system
1
1mM U-15N,13C nucleolin RBD12 in complex with unlabeled RNA
90% H2O/10% D2O
2
1mM U-15N nucleolin RBD12 in complex with unlabeled RNA
90% H2O/10% D2O
3
1mM U-15N nucleolin RBD12 in complex with unlabeled RNA
100% D2O
4
1mM U-15N-13C RNA in complex with U-15N nucleolin RBD12
100% D2O
Sample conditions
Conditions-ID
Ionic strength
pH
Pressure (kPa)
Temperature (K)
1
150
6.2
ambient
303K
2
150
6.2
ambient
318K
Crystal grow
*PLUS
Method: other / Details: NMR
-
NMR measurement
NMR spectrometer
Type
Manufacturer
Model
Field strength (MHz)
Spectrometer-ID
Bruker DRX
Bruker
DRX
600
1
Bruker DRX
Bruker
DRX
600
2
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Processing
NMR software
Name
Version
Developer
Classification
X-PLOR
3.841
BRUNGER
refinement
X-PLOR
3.841
BRUNGER
structuresolution
Refinement
Method: simulated annealing using XPLOR 3.841 / Software ordinal: 1 Details: structures are based on 3246 constraints, 3010 are NOE-derived including 150 intermolecular
NMR representative
Selection criteria: lowest energy
NMR ensemble
Conformer selection criteria: all calculated structures submitted Conformers calculated total number: 40 / Conformers submitted total number: 19
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