BMRB Entry 16813

Assembly:

Entities:

Entity 1, Ubl 81 residues - 8738.083 Da.

Residues 1-5 represent a non-native tag generated after 3C protease cleavage of N-terminal GST

Entity 2, SH3 71 residues - 7121.919 Da.

Residues 1-9 represent a non-native tag generated after thrombin protease cleavage of N-terminal GST

Samples:

Ubl_SH3_D2O: Ubl, [U-99% 13C; U-99% 15N], 1.2 ± 0.1 mM; SH3 1.2 ± 0.1 mM; D2O 100%; HEPES 10 mM; NaCl 50 mM; DTT 1 mM; EDTA 0.1 mM

SH3_Ubl_D2O: SH3, [U-99% 13C; U-99% 15N], 1.2 mM; Ubl 1.2 mM; D2O 100%; HEPES 10 mM; NaCl 50 mM; DTT 1 mM; EDTA 0.1 mM

Ubl_SH3_H2O: Ubl, [U-99% 13C; U-99% 15N], 0.8 mM; SH3 1.6 mM; D2O 5%; H2O 95%; HEPES 10 mM; NaCl 50 mM; DTT 1 mM; EDTA 0.1 mM

SH3_Ubl_H2O: SH3, [U-99% 13C; U-99% 15N], 0.8 mM; Ubl 1.6 mM; D2O 5%; H2O 95%; HEPES 10 mM; NaCl 50 mM; DTT 1 mM; EDTA 0.1 mM

Ubl_SH3_H2OB: Ubl, [U-99% 15N], 0.8 mM; SH3 1.6 mM; D2O 5%; H2O 95%; polyacrylamide 4.5%; HEPES 10 mM; NaCl 50 mM; DTT 1 mM; EDTA 0.1 mM

SH3_Ubl_H2OB: SH3, [U-99% 15N], 0.8 mM; Ubl 1.6 mM; D2O 5%; H2O 95%; polyacrylamide 4.5%; HEPES 10 mM; NaCl 50 mM; DTT 1 mM; EDTA 0.1 mM

isotropic: ionic strength: 50 mM; pH: 7.0; pressure: 1 atm; temperature: 303 K

polyacrylamide: ionic strength: 50 mM; pH: 7.0; pressure: 1 atm; temperature: 303 K

Experiments:

Software:

NMRPipe v2.5, Delaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax - processing

SPARKY v3.115, Goddard - chemical shift assignment, peak picking

CNS v1.2, Brunger, Adams, Clore, Gros, Nilges and Read - refinement, structure solution

NMR spectrometers:

• Bruker Avance 600 MHz
• Varian INOVA 800 MHz