ISDA: 2DQ5

SOLUTION NMR

NMR Experiment

Experiment	Type	Sample Contents	Ionic Strength	Solvent	pH	Pressure	Temprature (K)
1	15N-H1 HSQC	0.75-1.0mM 15N & 15N/13C labeled BBOX2; 50mM Tris-HCL, 100mM NaCl, 1mM ZnCl2, 10mM beta-mercaptoethanol; 2% sodium azide; 90% H2O, 10% D2O	50mM Tris-HCl; 100mM NaCl; 5mM ZnCl2, 10mM beta-mercaptoethanol	90% H2O/10% D2O	7.5	ambient	294
2	HNCA and HNCACB	0.75-1.0mM 15N & 15N/13C labeled BBOX2; 50mM Tris-HCL, 100mM NaCl, 1mM ZnCl2, 10mM beta-mercaptoethanol; 2% sodium azide; 90% H2O, 10% D2O	50mM Tris-HCl; 100mM NaCl; 5mM ZnCl2, 10mM beta-mercaptoethanol	90% H2O/10% D2O	7.5	ambient	294
3	3D-1H-15N-15N-HSQC-NOESY-HSQC	0.75-1.0mM 15N & 15N/13C labeled BBOX2; 50mM Tris-HCL, 100mM NaCl, 1mM ZnCl2, 10mM beta-mercaptoethanol; 2% sodium azide; 90% H2O, 10% D2O	50mM Tris-HCl; 100mM NaCl; 5mM ZnCl2, 10mM beta-mercaptoethanol	90% H2O/10% D2O	7.5	ambient	294
4	1H-15N-NOESY-HSQC and 1H-13C-NOESY-HSQC	0.75-1.0mM 15N & 15N/13C labeled BBOX2; 50mM Tris-HCL, 100mM NaCl, 1mM ZnCl2, 10mM beta-mercaptoethanol; 2% sodium azide; 90% H2O, 10% D2O	50mM Tris-HCl; 100mM NaCl; 5mM ZnCl2, 10mM beta-mercaptoethanol	90% H2O/10% D2O	7.5	ambient	294
5	15N-1H HSQC	0.75-1.0mM 15N & 15N/13C labeled BBOX2; 50mM Tris-HCL, 100mM NaCl, 1mM ZnCl2, 10mM beta-mercaptoethanol; 2% sodium azide; 90% H2O, 10% D2O	50mM Tris-HCl; 100mM NaCl; 5mM ZnCl2, 10mM beta-mercaptoethanol	90% H2O/10% D2O	7.5	ambient	294
6	1H-13C-HCCH-TOCSY	0.75-1.0mM 15N & 15N/13C labeled BBOX2; 50mM Tris-HCL, 100mM NaCl, 1mM ZnCl2, 10mM beta-mercaptoethanol; 2% sodium azide; 90% H2O, 10% D2O	50mM Tris-HCl; 100mM NaCl; 5mM ZnCl2, 10mM beta-mercaptoethanol	90% H2O/10% D2O	7.5	ambient	294

NMR Spectrometer Information

Spectrometer	Manufacturer	Model	Field Strength
1	Varian	INOVA	600

NMR Refinement

Method	Details	Software
torsion angle dynamics, low target function	A total of 200 random structures were calculated with CYANA2.1 using a fast torsion angle dynamics algorithm and the best structures were selected based on their low target function (less 1), no NOE violation less than 0.2 Ang, vdw < 0.25, dihedral <3o.	1

NMR Ensemble Information

Conformer Selection Criteria	structures with the lowest energy
Conformers Calculated Total Number	200
Conformers Submitted Total Number	16
Representative Model	1 (minimized average structure, lowest energy, fewest violations, closest to the average)

Computation: NMR Software

#	Classification	Version	Software Name	Author
1	processing	?	NMRPipe	Delaglio, F., Grzesiek, S., Vuister, G.W., Zhu, G. Pfeifer, J., Bax, A.
2	data analysis	3.110	Sparky	Goddard, T.D., Kneller, D.G.
3	data analysis	?	TALOS	Cornilescu, G. Delaglio, F., Bax, A.
4	refinement	2.1	CYANA	Guentert, P.