Experiment: 2MI7

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	2D 1H-15N HSQC	0.95 mM [U-100% 13C; U-100% 15N] a3Y, 30 mM [U-100% 2H] sodium acetate, 30 mM sodium chloride, 0.25 mM DSS, 0.02 % (w/v) sodium azide	92% H2O/8% D2O	60	5.6	ambient	303
2	2D 1H-13C HSQC	0.95 mM [U-100% 13C; U-100% 15N] a3Y, 30 mM [U-100% 2H] sodium acetate, 30 mM sodium chloride, 0.25 mM DSS, 0.02 % (w/v) sodium azide	99.99% D2O	60	5.6	ambient	303
3	3D HNCO	0.95 mM [U-100% 13C; U-100% 15N] a3Y, 30 mM [U-100% 2H] sodium acetate, 30 mM sodium chloride, 0.25 mM DSS, 0.02 % (w/v) sodium azide	92% H2O/8% D2O	60	5.6	ambient	303
4	3D HN(CA)CO	0.95 mM [U-100% 13C; U-100% 15N] a3Y, 30 mM [U-100% 2H] sodium acetate, 30 mM sodium chloride, 0.25 mM DSS, 0.02 % (w/v) sodium azide	92% H2O/8% D2O	60	5.6	ambient	303
5	3D CBCA(CO)NH	0.95 mM [U-100% 13C; U-100% 15N] a3Y, 30 mM [U-100% 2H] sodium acetate, 30 mM sodium chloride, 0.25 mM DSS, 0.02 % (w/v) sodium azide	92% H2O/8% D2O	60	5.6	ambient	303
6	3D HNCACB	0.95 mM [U-100% 13C; U-100% 15N] a3Y, 30 mM [U-100% 2H] sodium acetate, 30 mM sodium chloride, 0.25 mM DSS, 0.02 % (w/v) sodium azide	92% H2O/8% D2O	60	5.6	ambient	303
7	3D H(CC)(CO)NH TOCSY	0.95 mM [U-100% 13C; U-100% 15N] a3Y, 30 mM [U-100% 2H] sodium acetate, 30 mM sodium chloride, 0.25 mM DSS, 0.02 % (w/v) sodium azide	92% H2O/8% D2O	60	5.6	ambient	303
8	3D CC(CO)NH TOCSY	0.95 mM [U-100% 13C; U-100% 15N] a3Y, 30 mM [U-100% 2H] sodium acetate, 30 mM sodium chloride, 0.25 mM DSS, 0.02 % (w/v) sodium azide	92% H2O/8% D2O	60	5.6	ambient	303
9	3D HCCH-TOCSY	0.95 mM [U-100% 13C; U-100% 15N] a3Y, 30 mM [U-100% 2H] sodium acetate, 30 mM sodium chloride, 0.25 mM DSS, 0.02 % (w/v) sodium azide	99.99% D2O	60	5.6	ambient	303
10	3D (H)CCH3-TOCSY	0.95 mM [U-100% 13C; U-100% 15N] a3Y, 30 mM [U-100% 2H] sodium acetate, 30 mM sodium chloride, 0.25 mM DSS, 0.02 % (w/v) sodium azide	99.99% D2O	60	5.6	ambient	303
11	2D 1H-1H NOESY aromatic	0.95 mM [U-100% 13C; U-100% 15N] a3Y, 30 mM [U-100% 2H] sodium acetate, 30 mM sodium chloride, 0.25 mM DSS, 0.02 % (w/v) sodium azide	99.99% D2O	60	5.6	ambient	303
12	3D 1H-13C NOESY aromatic	0.95 mM [U-100% 13C; U-100% 15N] a3Y, 30 mM [U-100% 2H] sodium acetate, 30 mM sodium chloride, 0.25 mM DSS, 0.02 % (w/v) sodium azide	99.99% D2O	60	5.6	ambient	303
13	3D 1H-15N NOESY	0.95 mM [U-100% 13C; U-100% 15N] a3Y, 30 mM [U-100% 2H] sodium acetate, 30 mM sodium chloride, 0.25 mM DSS, 0.02 % (w/v) sodium azide	92% H2O/8% D2O	60	5.6	ambient	303
14	4D 1H-15N NOESY	0.95 mM [U-100% 13C; U-100% 15N] a3Y, 30 mM [U-100% 2H] sodium acetate, 30 mM sodium chloride, 0.25 mM DSS, 0.02 % (w/v) sodium azide	92% H2O/8% D2O	60	5.6	ambient	303
15	4D 1H-13C NOESY	0.95 mM [U-100% 13C; U-100% 15N] a3Y, 30 mM [U-100% 2H] sodium acetate, 30 mM sodium chloride, 0.25 mM DSS, 0.02 % (w/v) sodium azide	99.99% D2O	60	5.6	ambient	303
16	2D 1H-13C HSQC	0.95 mM [U-10% 13C; U-100% 15N] a3Y, 30 mM [U-100% 2H] sodium acetate, 30 mM sodium chloride, 0.25 mM DSS, 0.02 % (w/v) sodium azide	99.99% D2O	60	5.6	ambient	303
17	2D 1H-13C HSQC	0.95 mM [U-100% 13C; U-100% 15N] a3Y, 30 mM [U-100% 2H] sodium acetate, 30 mM sodium chloride, 0.25 mM DSS, 0.02 % (w/v) sodium azide	99.99% D2O	60	5.6	ambient	303

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Bruker	AVANCE	750
2	Bruker	AVANCE	500

NMR Refinement
Method	Details	Software
simulated annealing		TopSpin

NMR Ensemble Information
Conformer Selection Criteria	NOE violation cutoff >0.1A, bb dihedral angle violation cutoff >2 degrees, then lowest energy
Conformers Calculated Total Number	1000
Conformers Submitted Total Number	32
Representative Model	1 (lowest energy)

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	collection	TopSpin		Bruker Biospin
2	processing	Felix		Accelrys Software Inc.
3	chemical shift assignment	Sparky	3.113	Goddard
4	peak picking	Sparky	3.113	Goddard
5	noe integration	Sparky	3.113	Goddard
6	secondary structure analysis	TALOS+		Cornilescu, Delaglio and Bax
7	structure calculation	CNS	1.21	Brunger, Adams, Clore, Gros, Nilges and Read
8	refinement	CNS	1.21	Brunger, Adams, Clore, Gros, Nilges and Read
9	structure evaluation	PSVS	1.5	Bhattacharya and Montelione

RCSB PDB Core Operations are funded by the U.S. National Science Foundation (DBI-2321666), 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.