1DBY

NMR STRUCTURES OF CHLOROPLAST THIOREDOXIN M CH2 FROM THE GREEN ALGA CHLAMYDOMONAS REINHARDTII

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	3D_13C-SEPARATED_NOESY	2-3 MM THIOREDOXIN M (OXIDIZED) U-15N; 100 MM POTASSIUM PHOSPHATE BUFFER PH 5.8; 90% H2O 10% D20		100 mM POTASSIUM PHOSPHATE	5.8	AMBIENT	311
2	3D-SEPARATED-TOCSY	2-3 MM THIOREDOXIN M (OXIDIZED) U-15N; 100 MM POTASSIUM PHOSPHATE BUFFER PH 5.8; 90% H2O 10% D20		100 mM POTASSIUM PHOSPHATE	5.8	AMBIENT	311
3	2D NOESY	2-3 MM THIOREDOXIN M (OXIDIZED);100 MM POTASSIUM PHOSPHATE BUFFER PH 5.8; 90% H2O 10% D20		100 mM POTASSIUM PHOSPHATE	5.8	AMBIENT	311
4	2D NOESY	2-3 MM THIOREDOXIN M (OXIDIZED);100 MM POTASSIUM PHOSPHATE BUFFER PH 5.8; 100% D20		100 mM POTASSIUM PHOSPHATE	5.8	AMBIENT	311
5	TOCSY	2-3 MM THIOREDOXIN M (OXIDIZED);100 MM POTASSIUM PHOSPHATE BUFFER PH 5.8; 90% H2O 10% D20		100 mM POTASSIUM PHOSPHATE	5.8	AMBIENT	311

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Bruker	AMX	600
2	Bruker	DRX	500

NMR Refinement
Method	Details	Software
SIMULATED ANNEALING STARTING FROM A RANDOM ARRAY OF ATOMS. HIGH TEMPERATURE SIMULATED ANNEALING. RESTRAINED MOLECULAR DYNAMIC AT ROOM TEMPERATURE.	THE STRUCTURES ARE BASED ON 1372 TOTAL INTERPROTON DISTANCE RESTRAINTS, 1348 ARE DERIVED FROM NOE DATA AND 44 DERIVED FROM SLOW DEUTERIUM EXCHANGE EXPERIMENTS OF HYDROGEN BONDED AMIDE PROTONS IN REGULAR SECONDARY STRUCTURE MOTIFS. THE RESTRAINT SET INCLUDES 102 DIHEDRAL RESTRAINTS, 66 PHI, 34 KHI1, 2 KHI2. FINAL STRUCTURES ARE REFINED AGAINST THE FULL DESCRIPTION OF AMBER4 FORCE FIELD WITH REDUCED CHARGES FOR FORMALLY CHARGED GROUPS AS ARG, LYS, ASP, GLU RESIDUES.	XwinNMR

NMR Ensemble Information
Conformer Selection Criteria	STRUCTURES WITH ACCEPTABLE COVALENT GEOMETRY. STRUCTURES WITH FAVORABLE NON- BOND ENERGY. STRUCTURES WITH THE LEAST RESTRAINT VIOLATIONS. STRUCTURES WITH THE LOWEST ENERGY.
Conformers Calculated Total Number	50
Conformers Submitted Total Number	28
Representative Model	16 (closest to the average)

Additional NMR Experimental Information
Details	THE STRUCTURE WAS DETERMINED USING 2D HOMONUCLEAR IN H20 AND 100% D2O AND 3D- 15N SEPARATED NOESY AND TOCSY DATA.

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	collection	XwinNMR	2.0	BRUKER SPECTROSPIN AG
2	data analysis	Felix	2.3	BIOSYM TECHNOLOGIES
3	processing	NMRPipe	1.7	DELAGLIO
4	structure solution	Discover	2.3.0	BIOSYM TECHNOLOGIES
5	collection	UXNMR	940501.3	BRUKER SPECTROSPIN AG
6	processing	Gifa	4.0	DELSUC
7	refinement	Discover	2.3.0	BIOSYM TECHNOLOGIES

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.