2eu0 Experiments and Validation

Solution NMR

Source organism: Mus musculus [10090]
Refinement method: Distance geometry simulated annealing was used for refinement
Number of deposited models: 20 models

Sample information

Author description:
The NMR ensemble structure of the Itk SH2 domain bound to a phosphopeptide
Source organism: Mus musculus [10090]
Expression systems:
  • Not provided
  • Escherichia coli BL21(DE3) [469008]

Validation information

 
Structure ensemble information
Refinement method: Distance geometry simulated annealing was used for refinement
Number of calculated models: 200
Number of deposited models: 20 (structures with the least restraint violations)
Representative model: 1 (lowest energy)

Experimental information

Sample 1
Contents: 1mM Itk SH2 domain, 20mM phosphopeptide, 50mM KH2PO4, 75mM NaCl, 2mM DTT, 0.02% NaN3, pH 7.4 Recorded spectra: 2D NOESY

Sample 2
Contents: U-15N labeled, 1mM Itk SH2 domain, 20mM phosphopeptide, 50mM KH2PO4, 75mM NaCl, 2mM DTT, 0.02% NaN3, pH 7.4 Recorded spectra: 3D_15N-separated_NOESY, HNHA

Sample 3
Contents: U-15N, 13-C labeled, 1mM Itk SH2 domain, 20mM phosphopeptide, 50mM KH2PO4, 75mM NaCl, 2mM DTT, 0.02% NaN3, pH 7.4 Recorded spectra: 13C edited_13C/15_N filtered NOESY, 15N_edited_13C/15N_filtered NOESY, 3D_13C-separated_NOESY, 3D_13C_separated_aromatic_NOESY, CBCA(CO)NH, CCONH, HCCH_TOCSY, HNCACB

Sample 4
Contents: 5mM phosphopeptide, 50mM KH2PO4, 75mM NaCl, 2mM DTT, 0.02% NaN3, pH 7.4 Recorded spectra: 2D TOCSY

Sample 5
Contents: 1mM Itk SH2 domain, 20mM phosphopeptide, 50mM KH2PO4, 75mM NaCl, 2mM DTT, 0.02% NaN3, pH 7.4 Recorded spectra: D2O exchange experiment

Samples and spectra
NMR spectrometers used
Bruker DRX 500 MHz