6AMR

Solution NMR structure of a putative thioredoxin (ECH_0218) in the reduced state from Ehrlichia chaffeensis, the etiological agent responsible for human monocytic ehrlichiosis. Seattle Structural Genomics Center for Infectious Disease target EhchA.00546.a

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	2D 1H-15N HSQC	100 mM sodium chloride, 20 mM TRIS, 1 mM DTT	93% H2O/7% D2O	0.12 M	7	1 atm	293	Varian VXRS 800
2	2D 1H-13C HSQC aliphatic	100 mM sodium chloride, 20 mM TRIS, 1 mM DTT	93% H2O/7% D2O	0.12 M	7	1 atm	293	Varian VXRS 800
3	2D 1H-13C HSQC aromatic	100 mM sodium chloride, 20 mM TRIS, 1 mM DTT	93% H2O/7% D2O	0.12 M	7	1 atm	293	Varian VXRS 800
4	3D C(CO)NH	100 mM sodium chloride, 20 mM TRIS, 1 mM DTT	93% H2O/7% D2O	0.12 M	7	1 atm	293	Varian VXRS 600
5	3D HNCACB	100 mM sodium chloride, 20 mM TRIS, 1 mM DTT	93% H2O/7% D2O	0.12 M	7	1 atm	293	Varian VXRS 600
11	3D HNCO	100 mM sodium chloride, 20 mM TRIS, 1 mM DTT	93% H2O/7% D2O	0.12 M	7	1 atm	293	Varian VXRS 600
10	3D 1H-15N NOESY	100 mM sodium chloride, 20 mM TRIS, 1 mM DTT	93% H2O/7% D2O	0.12 M	7	1 atm	293	Varian VXRS 800
9	3D 1H-13C NOESY aliphatic	100 mM sodium chloride, 20 mM TRIS, 1 mM DTT	93% H2O/7% D2O	0.12 M	7	1 atm	293	Varian VXRS 750
8	3D 1H-13C NOESY aromatic	100 mM sodium chloride, 20 mM TRIS, 1 mM DTT	93% H2O/7% D2O	0.12 M	7	1 atm	293	Varian VXRS 750
7	2D 1H-15N HSQC Deuterium Exchange	100 mM sodium chloride, 20 mM TRIS, 1 mM DTT	93% H2O/7% D2O	0.12 M	7	1 atm	293	Varian VXRS 750

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Varian	VXRS	600
2	Varian	VXRS	750
3	Varian	VXRS	800

NMR Refinement
Method	Details	Software
torsion angle dynamics	Proline-97 was constrained to the Cis configuration on the basis of other Trx structures. STRUCTURE DETERMINATION WAS PERFORMED ITERATIVELY USING CYANA (AUTOMATED NOESY ASSIGNMENTS). A TOTAL OF 20 STRUCTURES OUT OF 100 WITH LOWEST TARGET FUNCTION FROM THE FINAL CYANA CALCULATION WERE TAKEN AND REFINED BY RESTRAINED MOLECULAR DYNAMICS/ENERGY MINIMIZATION IN EXPLICIT WATER (CNS) AFTER ADDING 10% TO THE UPPER BOUNDARY LIMIT OF THE DISTANCE RESTRAINTS AND THE VDW LIMIT TO THE LOWER RESTRAINT. PARAM19 WAS USED FOR THE WATER REFINEMENT CALCULATIONS.	CYANA

NMR Ensemble Information
Conformer Selection Criteria	target function
Conformers Calculated Total Number	100
Conformers Submitted Total Number	20
Representative Model	1 (closest to the average)

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	refinement	CNS	1.1	Brunger A. T. et.al.
2	data analysis	Sparky	3.115	Goddard
3	data analysis	PSVS	1.5	Bhattacharya and Montelione
4	refinement	CYANA	2.1	Guntert, Mumenthaler and Wuthrich
5	data analysis	TALOS	+	Cornilescu, Delaglio and Bax
6	processing	Felix	2007	Accelrys Software Inc.
7	structure calculation	CYANA	2.1	Guntert, Mumenthaler and Wuthrich

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.