6TR8

Corynebacterium diphtheriae methionine sulfoxide reductase B (MsrB) solution structure - reduced form

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	2D 1H-15N HSQC	1 mM [U-100% 13C; U-100% 15N] MsrB, 20 mM sodium phosphate, 150 mM sodium chloride, 2 mM DTT, 10 % [U-100% 2H] D2O	90% H2O/10% D2O	165 mM	7.0	1 atm	298	Bruker AVANCE 800
2	3D HNCACB	1 mM [U-100% 13C; U-100% 15N] MsrB, 20 mM sodium phosphate, 150 mM sodium chloride, 2 mM DTT, 10 % [U-100% 2H] D2O	90% H2O/10% D2O	165 mM	7.0	1 atm	298	Bruker AVANCE 800
3	3D HN(COCA)CB	1 mM [U-100% 13C; U-100% 15N] MsrB, 20 mM sodium phosphate, 150 mM sodium chloride, 2 mM DTT, 10 % [U-100% 2H] D2O	90% H2O/10% D2O	165 mM	7.0	1 atm	298	Bruker AVANCE 800
4	3D HNCO	1 mM [U-100% 13C; U-100% 15N] MsrB, 20 mM sodium phosphate, 150 mM sodium chloride, 2 mM DTT, 10 % [U-100% 2H] D2O	90% H2O/10% D2O	165 mM	7.0	1 atm	298	Bruker AVANCE 800
5	3D HN(CA)CO	1 mM [U-100% 13C; U-100% 15N] MsrB, 20 mM sodium phosphate, 150 mM sodium chloride, 2 mM DTT, 10 % [U-100% 2H] D2O	90% H2O/10% D2O	165 mM	7.0	1 atm	298	Bruker AVANCE 800
6	3D HBHA(CO)NH	1 mM [U-100% 13C; U-100% 15N] MsrB, 20 mM sodium phosphate, 150 mM sodium chloride, 2 mM DTT, 10 % [U-100% 2H] D2O	90% H2O/10% D2O	165 mM	7.0	1 atm	298	Bruker AVANCE 800
7	3D HCCH-TOCSY	1 mM [U-100% 13C; U-100% 15N] MsrB, 20 mM sodium phosphate, 150 mM sodium chloride, 2 mM DTT, 10 % [U-100% 2H] D2O	90% H2O/10% D2O	165 mM	7.0	1 atm	298	Bruker AVANCE 800
8	3D HCCH-TOCSY	1 mM [U-100% 13C; U-100% 15N] MsrB, 20 mM sodium phosphate, 150 mM sodium chloride, 2 mM DTT, 10 % [U-100% 2H] D2O	90% H2O/10% D2O	165 mM	7.0	1 atm	298	Bruker AVANCE 800
9	2D 1H-13C HSQC	1 mM [U-100% 13C; U-100% 15N] MsrB, 20 mM sodium phosphate, 150 mM sodium chloride, 2 mM DTT, 10 % [U-100% 2H] D2O	90% H2O/10% D2O	165 mM	7.0	1 atm	298	Bruker AVANCE 800
10	2D 1H-13C HSQC aromatic	1 mM [U-100% 13C; U-100% 15N] MsrB, 20 mM sodium phosphate, 150 mM sodium chloride, 2 mM DTT, 10 % [U-100% 2H] D2O	90% H2O/10% D2O	165 mM	7.0	1 atm	298	Bruker AVANCE 800
11	3D 1H-15N NOESY	1 mM [U-100% 13C; U-100% 15N] MsrB, 20 mM sodium phosphate, 150 mM sodium chloride, 2 mM DTT, 10 % [U-100% 2H] D2O	90% H2O/10% D2O	165 mM	7.0	1 atm	298	Bruker AVANCE 800
12	3D 1H-13C NOESY aliphatic	1 mM [U-100% 13C; U-100% 15N] MsrB, 20 mM sodium phosphate, 150 mM sodium chloride, 2 mM DTT, 10 % [U-100% 2H] D2O	90% H2O/10% D2O	165 mM	7.0	1 atm	298	Bruker AVANCE 800
13	3D 1H-13C NOESY aromatic	1 mM [U-100% 13C; U-100% 15N] MsrB, 20 mM sodium phosphate, 150 mM sodium chloride, 2 mM DTT, 10 % [U-100% 2H] D2O	90% H2O/10% D2O	165 mM	7.0	1 atm	298	Bruker AVANCE 800
14	2D (HB)CB(CGCD)HD	1 mM [U-100% 13C; U-100% 15N] MsrB, 20 mM sodium phosphate, 150 mM sodium chloride, 2 mM DTT, 10 % [U-100% 2H] D2O	90% H2O/10% D2O	165 mM	7.0	1 atm	298	Bruker AVANCE 800
15	2D (HB)CB(CGCDCE)HE	1 mM [U-100% 13C; U-100% 15N] MsrB, 20 mM sodium phosphate, 150 mM sodium chloride, 2 mM DTT, 10 % [U-100% 2H] D2O	90% H2O/10% D2O	165 mM	7.0	1 atm	298	Bruker AVANCE 800

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Bruker	AVANCE	800

NMR Refinement
Method	Details	Software
simulated annealing		CNS
torsion angle dynamics		X-PLOR NIH

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

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	collection	TopSpin	3.5	Bruker Biospin
2	processing	TopSpin	3.5	Bruker Biospin
3	processing	NMRPipe		Delaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax
4	data analysis	CcpNmr Analysis		CCPN
5	peak picking	CcpNmr Analysis		CCPN
6	chemical shift assignment	CcpNmr Analysis		CCPN
7	structure calculation	CYANA		Guntert, Mumenthaler and Wuthrich
8	refinement	CNS		Brunger, Adams, Clore, Gros, Nilges and Read
9	refinement	X-PLOR NIH		Schwieters, Kuszewski, Tjandra and Clore

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.