Growthfactorreceptor-boundprotein2 / Adapter protein GRB2 / Protein Ash / SH2/SH3 adapter GRB2
Mass: 11204.756 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Homo sapiens (human) / Gene: GRB2, ASH / Production host: Escherichia coli BL21(DE3) (bacteria) / Strain (production host): BL21(DE3) / References: UniProt: P62993
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Experimental details
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Experiment
Experiment
Method: SOLUTION NMR
NMR experiment
Conditions-ID
Experiment-ID
Solution-ID
Sample state
Spectrometer-ID
Type
1
1
1
isotropic
1
2D 1H-15N HSQC
1
2
1
isotropic
1
3D 1H-13C NOESY aliphatic
1
3
1
isotropic
1
3D 1H-13C NOESY aromatic
1
4
1
isotropic
1
3D 1H-15N NOESY
1
5
1
isotropic
2
3D HN(CA)CB
1
6
1
isotropic
3
3D HNCA
1
7
1
isotropic
2
3DCBCA(CO)NH
1
8
1
isotropic
2
3DHBHA(CO)NH
1
9
1
isotropic
2
3D HNCO
1
10
1
isotropic
2
3DH(CCO)NH
1
11
1
isotropic
2
3D (H)CCH-TOCSY
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Sample preparation
Details
Type: solution Contents: 200 mM sodium chloride, 20 mM sodium phosphate, 1 mM EDTA, 1 mM PMSF, 5 mM sodium azide, 300 uM [U-100% 13C; U-100% 15N] Grb2-SH2, 90% H2O/10% D2O Details: ~300 uM 13C/15N-Grb2-SH2 in buffer 20 mM NaPi pH 7.0 containing 200 mM NaCl, 1.0 mM EDTA, 1.0 mM PMSF, 5.0 mM NaN3, and 10% D2O Label: Grb2-SH2 / Solvent system: 90% H2O/10% D2O
Sample
Conc. (mg/ml)
Component
Isotopic labeling
Solution-ID
200mM
sodiumchloride
naturalabundance
1
20mM
sodiumphosphate
naturalabundance
1
1mM
EDTA
naturalabundance
1
1mM
PMSF
naturalabundance
1
5mM
sodiumazide
naturalabundance
1
300uM
Grb2-SH2
[U-100% 13C; U-100% 15N]
1
Sample conditions
Details: ~300 uM 13C/15N-Grb2-SH2 in buffer 20 mM NaPi pH 7.0 containing 200 mM NaCl, 1.0 mM EDTA, 1.0 mM PMSF, 5.0 mM NaN3, and 10% D2O Ionic strength: 138.2 mM / Label: Condition1 / pH: 7 / Pressure: 1 atm / Temperature: 298 K
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NMR measurement
NMR spectrometer
Type
Manufacturer
Model
Field strength (MHz)
Spectrometer-ID
Bruker AVANCE III HD
Bruker
AVANCEIIIHD
900
1
Bruker AVANCE III
Bruker
AVANCEIII
800
2
Bruker AVANCE III HD
Bruker
AVANCEIIIHD
700
3
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Processing
NMR software
Name
Version
Developer
Classification
CS-ROSETTA
3.3
Shen, Vernon, BakerandBax
structurecalculation
CcpNmr Analysis
CCPN
dataanalysis
ARIA
Linge, O'DonoghueandNilges
structurecalculation
CS-ROSETTA
Shen, Vernon, BakerandBax
refinement
CcpNmr Analysis
CCPN
chemicalshiftassignment
CcpNmr Analysis
CCPN
peakpicking
Refinement
Method: na / Software ordinal: 4
NMR representative
Selection criteria: lowest energy
NMR ensemble
Conformer selection criteria: structures with the lowest energy Conformers calculated total number: 66317 / Conformers submitted total number: 20
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