2K5T

Solution NMR Structure of Putative N-Acetyl Transferase YhhK from E. coli Bound to Coenzyme A: Northeast Structural Genomics Consortium Target ET106

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	2D 1H-15N HSQC	0.6 mM [U-100% 13C; U-100% 15N] YhhK, 5 mM Coenzyme A, 7 % D2O, 300 mM sodium chloride, 25 mM TRIS, 5 mM DTT	93% H2O/7% D2O	300	7	ambient	293
2	2D 1H-13C HSQC	0.6 mM [U-100% 13C; U-100% 15N] YhhK, 5 mM Coenzyme A, 7 % D2O, 300 mM sodium chloride, 25 mM TRIS, 5 mM DTT	93% H2O/7% D2O	300	7	ambient	293
3	3D CBCA(CO)NH	0.6 mM [U-100% 13C; U-100% 15N] YhhK, 5 mM Coenzyme A, 7 % D2O, 300 mM sodium chloride, 25 mM TRIS, 5 mM DTT	93% H2O/7% D2O	300	7	ambient	293
4	3D HNCACB	0.6 mM [U-100% 13C; U-100% 15N] YhhK, 5 mM Coenzyme A, 7 % D2O, 300 mM sodium chloride, 25 mM TRIS, 5 mM DTT	93% H2O/7% D2O	300	7	ambient	293
5	3D HNCO	0.6 mM [U-100% 13C; U-100% 15N] YhhK, 5 mM Coenzyme A, 7 % D2O, 300 mM sodium chloride, 25 mM TRIS, 5 mM DTT	93% H2O/7% D2O	300	7	ambient	293
6	3D HCCH-TOCSY	0.6 mM [U-100% 13C; U-100% 15N] YhhK, 5 mM Coenzyme A, 7 % D2O, 300 mM sodium chloride, 25 mM TRIS, 5 mM DTT	93% H2O/7% D2O	300	7	ambient	293
7	3D HCCH-COSY	0.6 mM [U-100% 13C; U-100% 15N] YhhK, 5 mM Coenzyme A, 7 % D2O, 300 mM sodium chloride, 25 mM TRIS, 5 mM DTT	93% H2O/7% D2O	300	7	ambient	293
8	3D HBHA(CO)NH	0.6 mM [U-100% 13C; U-100% 15N] YhhK, 5 mM Coenzyme A, 7 % D2O, 300 mM sodium chloride, 25 mM TRIS, 5 mM DTT	93% H2O/7% D2O	300	7	ambient	293
9	3D 1H-15N NOESY	0.6 mM [U-100% 13C; U-100% 15N] YhhK, 5 mM Coenzyme A, 7 % D2O, 300 mM sodium chloride, 25 mM TRIS, 5 mM DTT	93% H2O/7% D2O	300	7	ambient	293
10	3D 1H-13C NOESY	0.6 mM [U-100% 13C; U-100% 15N] YhhK, 5 mM Coenzyme A, 7 % D2O, 300 mM sodium chloride, 25 mM TRIS, 5 mM DTT	93% H2O/7% D2O	300	7	ambient	293
11	4D 13C HMQC-NOESY-HMQC	0.6 mM [U-100% 13C; U-100% 15N] YhhK, 5 mM Coenzyme A, 100 % D2O, 300 mM sodium chloride, 25 mM TRIS, 5 mM DTT	100% D2O	300	7	ambient	293

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Varian	INOVA	750
2	Varian	INOVA	600

NMR Refinement
Method	Details	Software
molecular dynamics, simulated annealing		Felix

NMR Ensemble Information
Conformer Selection Criteria	fewest restraints violations and energies, favorable backbone geometry, minimal clashes
Conformers Calculated Total Number	30
Conformers Submitted Total Number	20
Representative Model	1 (no criterion)

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	processing	Felix		Accelrys Software Inc.
2	chemical shift assignment	Felix		Accelrys Software Inc.
3	data analysis	Felix		Accelrys Software Inc.
4	processing	Sparky		Goddard
5	chemical shift assignment	Sparky		Goddard
6	data analysis	Sparky		Goddard
7	structure solution	AutoStructure		Huang, Tejero, Powers and Montelione
8	refinement	AutoStructure		Huang, Tejero, Powers and Montelione
9	structure solution	CNS		Brunger, Adams, Clore, Gros, Nilges and Read
10	refinement	CNS		Brunger, Adams, Clore, Gros, Nilges and Read
11	structure solution	X-PLOR NIH		Schwieters, Kuszewski, Tjandra and Clore
12	refinement	X-PLOR NIH		Schwieters, Kuszewski, Tjandra and Clore

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.