Experiment: 5MTI

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	2D 1H-1H NOESY	0.5 mM [U-99% 15N] 15N_PCP17, 50 mM potassium phosphate, 200 mM sodium chloride, 1 % DSS, 90 % H2O, 10 % D2O	90% H2O/10% D2O	250 mM	6.50	1 atm	288	Bruker AVANCE II 700
2	3D 1H-15N NOESY	0.5 mM [U-99% 15N] 15N_PCP17, 50 mM potassium phosphate, 200 mM sodium chloride, 1 % DSS, 90 % H2O, 10 % D2O	90% H2O/10% D2O	250 mM	6.50	1 atm	288	Bruker AVANCE II 700
3	3D 1H-13C NOESY	0.3 mM [U-13C; U-15N] 13C,15N_PCP17, 50 mM potassium phosphate, 200 mM sodium chloride, 1 % DSS, 90 % H2O, 10 % D2O	90% H2O/10% D2O	250 mM	6.50	1 atm	288	Bruker AVANCE II 700
4	3D HNCO	0.3 mM [U-13C; U-15N] 13C,15N_PCP17, 50 mM potassium phosphate, 200 mM sodium chloride, 1 % DSS, 90 % H2O, 10 % D2O	90% H2O/10% D2O	250 mM	6.50	1 atm	288	Bruker AVANCE II 700
5	3D HN(CA)CO	0.3 mM [U-13C; U-15N] 13C,15N_PCP17, 50 mM potassium phosphate, 200 mM sodium chloride, 1 % DSS, 90 % H2O, 10 % D2O	90% H2O/10% D2O	250 mM	6.50	1 atm	288	Bruker AVANCE II 700
6	3D HNCA	0.3 mM [U-13C; U-15N] 13C,15N_PCP17, 50 mM potassium phosphate, 200 mM sodium chloride, 1 % DSS, 90 % H2O, 10 % D2O	90% H2O/10% D2O	250 mM	6.50	1 atm	288	Bruker AVANCE II 700
7	3D HN(CO)CA	0.3 mM [U-13C; U-15N] 13C,15N_PCP17, 50 mM potassium phosphate, 200 mM sodium chloride, 1 % DSS, 90 % H2O, 10 % D2O	90% H2O/10% D2O	250 mM	6.50	1 atm	288	Bruker AVANCE II 700
8	3D HNCACB	0.3 mM [U-13C; U-15N] 13C,15N_PCP17, 50 mM potassium phosphate, 200 mM sodium chloride, 1 % DSS, 90 % H2O, 10 % D2O	90% H2O/10% D2O	250 mM	6.50	1 atm	288	Bruker AVANCE II 700
9	3D HN(COCA)CB	0.3 mM [U-13C; U-15N] 13C,15N_PCP17, 50 mM potassium phosphate, 200 mM sodium chloride, 1 % DSS, 90 % H2O, 10 % D2O	90% H2O/10% D2O	250 mM	6.50	1 atm	288	Bruker AVANCE II 700
10	3D HBHA(CO)NH	0.3 mM [U-13C; U-15N] 13C,15N_PCP17, 50 mM potassium phosphate, 200 mM sodium chloride, 1 % DSS, 90 % H2O, 10 % D2O	90% H2O/10% D2O	250 mM	6.50	1 atm	288	Bruker AVANCE II 700
11	3D HCCH-TOCSY	0.3 mM [U-13C; U-15N] 13C,15N_PCP17, 50 mM potassium phosphate, 200 mM sodium chloride, 1 % DSS, 90 % H2O, 10 % D2O	90% H2O/10% D2O	250 mM	6.50	1 atm	288	Bruker AVANCE II 700
12	2D 1H-15N HSQC	0.5 mM [U-99% 15N] 15N_PCP17, 50 mM potassium phosphate, 200 mM sodium chloride, 1 % DSS, 90 % H2O, 10 % D2O	90% H2O/10% D2O	250 mM	6.50	1 atm	288	Bruker AVANCE II 700
13	2D 1H-13C HSQC	0.3 mM [U-13C; U-15N] 13C,15N_PCP17, 50 mM potassium phosphate, 200 mM sodium chloride, 1 % DSS, 90 % H2O, 10 % D2O	90% H2O/10% D2O	250 mM	6.50	1 atm	288	Bruker AVANCE II 700

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Bruker	AVANCE II	700

NMR Refinement
Method	Details	Software
molecular dynamics	The 20 conformers of apo-PCP17 with the lowest target function values were subjected to restrained energy minimization and Molecular Dynamics (MD) calculations in explicit solvent with AMBER 14.0. NOEs and torsion angle constraints were applied with force constants of 50 kcal mol?1??2 and 32 kcal mol?1 rad?2, respectively.	Amber

NMR Ensemble Information
Conformer Selection Criteria	structures with the lowest energy
Conformers Calculated Total Number	200
Conformers Submitted Total Number	20
Representative Model	1 (target function)

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	collection	TopSpin	3.5pl6	Bruker Biospin
2	processing	TopSpin	3.5pl6	Bruker Biospin
3	data analysis	CARA	3	Keller and Wuthrich
4	chemical shift assignment	CARA	3	Keller and Wuthrich
5	peak picking	ATNOS/CANDID	UNIO	Herrmann, Guntert and Wuthrich
6	structure calculation	CYANA	2.1	Guntert, Mumenthaler and Wuthrich
7	refinement	Amber	14	Case, Darden, Cheatham III, Simmerling, Wang, Duke, Luo, ... and Kollman

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.