2L1P

NMR solution structure of the N-terminal domain of DNA-binding protein SATB1 from Homo sapiens: Northeast Structural Genomics Target HR4435B(179-250)

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	2D 1H-15N HSQC	1.08 mM [U-100% 13C; U-100% 15N] HR4435B	90% H2O/10% D2O	5mM CaCl2, 100mM NaCl	6.5	1 atm	298
2	2D 1H-13C HSQC	1.08 mM [U-100% 13C; U-100% 15N] HR4435B	90% H2O/10% D2O	5mM CaCl2, 100mM NaCl	6.5	1 atm	298
3	3D CBCA(CO)NH	1.08 mM [U-100% 13C; U-100% 15N] HR4435B	90% H2O/10% D2O	5mM CaCl2, 100mM NaCl	6.5	1 atm	298
4	3D HNCACB	1.08 mM [U-100% 13C; U-100% 15N] HR4435B	90% H2O/10% D2O	5mM CaCl2, 100mM NaCl	6.5	1 atm	298
5	3D HBHA(CO)NH	1.08 mM [U-100% 13C; U-100% 15N] HR4435B	90% H2O/10% D2O	5mM CaCl2, 100mM NaCl	6.5	1 atm	298
6	3D HNCO	1.08 mM [U-100% 13C; U-100% 15N] HR4435B	90% H2O/10% D2O	5mM CaCl2, 100mM NaCl	6.5	1 atm	298
7	3D HCCH-TOCSY	1.08 mM [U-100% 13C; U-100% 15N] HR4435B	90% H2O/10% D2O	5mM CaCl2, 100mM NaCl	6.5	1 atm	298
8	3D 1H-15N NOESY	1.08 mM [U-100% 13C; U-100% 15N] HR4435B	90% H2O/10% D2O	5mM CaCl2, 100mM NaCl	6.5	1 atm	298
9	3D 1H-13C NOESY	1.08 mM [U-100% 13C; U-100% 15N] HR4435B	90% H2O/10% D2O	5mM CaCl2, 100mM NaCl	6.5	1 atm	298
10	2D 1H-15N HSQC	1.08 mM [U-100% 13C; U-100% 15N] HR4435B	90% H2O/10% D2O	5mM CaCl2, 100mM NaCl	6.5	1 atm	298
11	2D 1H-13C HSQC	1.08 mM [U-100% 13C; U-100% 15N] HR4435B	90% H2O/10% D2O	5mM CaCl2, 100mM NaCl	6.5	1 atm	298

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Bruker	Avance	600
2	Bruker	Avance	800

NMR Refinement
Method	Details	Software
simulated annealing	The structure was determined using triple resonance NMR spectroscopy. Automated backbone assignments were made using Autoassign. Sidechain assignments were completed manually. Automated NOESY assignments were made using AutoStructure and structure solution was obtained using AutoStructure and CYANA-2.1. 150 structures were calculated and 20 best structures were refined in a shell of water using CNS. Initial dihedral angles were obtained using TALOS. The structure calculations were done including the N-terminal 6xHis tag. Resonance assignments were validated using AVS validation software. Final quality scores were determined using PSVS software. Ordered residues are defined as:11-13,18-54,62-77 . RMSD(ordered residuesall backbone aatoms 0.9A; All heavy atoms 1.4A; Ramachandran Statistics for all ordered residues: Most favoured 94.6%, additionally allowed region: 5.4%; Procheck scores for all ordered residues (Raw/Z) phi-psi 0.09/0.67; All dihedral angles 0.04/0.24; MolProbity clash score (Raw/Z) 13.00/-0.71. RPF scores for the goodness of fir of the structure to the NMR data: recall:0.933; Precision 0.918; F-measure 0.926 and final DP score: 0.768	AutoAssign

NMR Ensemble Information
Conformer Selection Criteria	structures with the lowest energy
Conformers Calculated Total Number	150
Conformers Submitted Total Number	20
Representative Model	1 (lowest energy)

Additional NMR Experimental Information
Details	Data acquisition on B600 was performed using 35 uL of 1.08mM sample and a 1.7mm microcryoprobe.

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	chemical shift assignment	AutoAssign	2.2.1	Zimmerman, Moseley, Kulikowski and Montelione
2	chemical shift assignment	AutoStructure	2.2.1	Huang, Tejero, Powers and Montelione
3	structure solution	AutoStructure	2.2.1	Huang, Tejero, Powers and Montelione
4	geometry optimization	CYANA	2.1	Guntert, Mumenthaler and Wuthrich
5	structure solution	CYANA	2.1	Guntert, Mumenthaler and Wuthrich
6	refinement	CNS	2.0.6	Brunger, Adams, Clore, Gros, Nilges and Read
7	geometry optimization	CNS	2.0.6	Brunger, Adams, Clore, Gros, Nilges and Read

RCSB PDB Core Operations are funded by the National Science Foundation (DBI-1832184), the US Department of Energy (DE-SC0019749), and the National Cancer Institute, National Institute of Allergy and Infectious Diseases, and National Institute of General Medical Sciences of the National Institutes of Health under grant R01GM133198.