Evidence: NMR Distance Restraints, distance restraints derived from NOESYs dihedral angle restraints derived from chemical shifts hydrogen bond restraints supported by 3j-HNCO
Type
Name
Symmetry operation
Number
identity operation
1_555
1
NMR ensembles
Data
Criteria
Number of conformers (submitted / calculated)
20 / 20
target function
Representative
Model #1
target function
-
Components
#1: Protein
Growthfactorreceptor-boundprotein2 / Downstream of receptor kinase / Protein enhancer of sevenless 2B
Mass: 6913.521 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Drosophila melanogaster (fruit fly) / Gene: drk / Production host: Escherichia coli BL21(DE3) (bacteria) / Strain (production host): BL21(DE3) / References: UniProt: Q08012
-
Experimental details
-
Experiment
Experiment
Method: SOLUTION NMR
NMR experiment
Conditions-ID
Experiment-ID
Solution-ID
Sample state
Spectrometer-ID
Type
1
1
1
isotropic
1
13C-separated NOESY
1
7
1
isotropic
1
15N-sperated NOESY
2
12
2
isotropic
1
2D 1H-15N HSQC
1
14
1
isotropic
1
3D HNCO
1
15
1
isotropic
1
3D HN(CA)CO
1
16
1
isotropic
1
3D HNCA
1
17
1
isotropic
1
3DHN(CO)CA
1
13
1
isotropic
1
3DCBCA(CO)NH
1
18
1
isotropic
1
3D CBCANH
1
19
1
isotropic
1
3D CC(CO)NH
1
20
1
isotropic
1
3DH(CCO)NH
1
21
1
isotropic
1
3D 1H-15N TOCSY
1
22
1
isotropic
1
3D 1H-15N COSY
1
23
1
isotropic
1
3D TROSYHNCO
-
Sample preparation
Details
Type
Solution-ID
Contents
Label
Solvent system
solution
1
1 mM [U-100% 13C; U-100% 15N] the C-terminal SH3 domain of Drosophila, 90% H2O/10% D2O
13C15N_Sample
90% H2O/10% D2O
solution
2
1 mM [U-100% 15N] the C-terminal SH3 domain of Drosophila, 90% H2O/10% D2O
15N_Sample
90% H2O/10% D2O
Sample
Conc. (mg/ml)
Component
Isotopic labeling
Solution-ID
1mM
the C-terminal SH3 domain of Drosophila
[U-100% 13C; U-100% 15N]
1
1mM
the C-terminal SH3 domain of Drosophila
[U-100% 15N]
2
Sample conditions
Conditions-ID
Ionic strength
Label
pH
Pressure (kPa)
Temperature (K)
1
100mM
13C15N_Sample
7.4
AMBIENTatm
298K
2
100mM
15N_Sample
7.4
AMBIENTatm
298K
-
NMR measurement
NMR spectrometer
Type: Bruker AVANCE III / Manufacturer: Bruker / Model: AVANCE III / Field strength: 600 MHz
-
Processing
NMR software
Name
Version
Developer
Classification
Azara
2.8
Boucher, W.
processing
CcpNmr Analysis
2.5.1
CCPN
chemicalshiftassignment
CYANA
3.98
Guntert, P., Mumenthaler, C., andWuthrich, K.
structurecalculation
TopSpin
3.5PL7
BrukerBiospin
collection
OPALp
Luginbuhl, R., Guntert, P., Billeter, M., andWuthrich, K.
refinement
Refinement
Method: molecular dynamics / Software ordinal: 5
NMR representative
Selection criteria: target function
NMR ensemble
Conformer selection criteria: target function / Conformers calculated total number: 20 / Conformers submitted total number: 20
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