2KF2

Solution NMR structure of of Streptomyces coelicolor polyketide cyclase SCO5315. Northeast Structural Genomics Consortium target RR365

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	2D 1H-15N HSQC	1 mM [U-99% 13C; U-99% 15N] entity-1, 20 mM MES-2, 100 mM sodium chloride-3, 5 mM calcium chloride-4, 10 mM DTT-5, 50 uM DSS-6, 0.02 % sodium azide-7	90% H2O/10% D2O	112	6.5	ambient	293
2	2D 1H-13C HSQC	1 mM [U-99% 13C; U-99% 15N] entity-1, 20 mM MES-2, 100 mM sodium chloride-3, 5 mM calcium chloride-4, 10 mM DTT-5, 50 uM DSS-6, 0.02 % sodium azide-7	90% H2O/10% D2O	112	6.5	ambient	293
3	3D CBCA(CO)NH	1 mM [U-99% 13C; U-99% 15N] entity-1, 20 mM MES-2, 100 mM sodium chloride-3, 5 mM calcium chloride-4, 10 mM DTT-5, 50 uM DSS-6, 0.02 % sodium azide-7	90% H2O/10% D2O	112	6.5	ambient	293
4	3D HNCACB	1 mM [U-99% 13C; U-99% 15N] entity-1, 20 mM MES-2, 100 mM sodium chloride-3, 5 mM calcium chloride-4, 10 mM DTT-5, 50 uM DSS-6, 0.02 % sodium azide-7	90% H2O/10% D2O	112	6.5	ambient	293
5	3D HBHA(CO)NH	1 mM [U-99% 13C; U-99% 15N] entity-1, 20 mM MES-2, 100 mM sodium chloride-3, 5 mM calcium chloride-4, 10 mM DTT-5, 50 uM DSS-6, 0.02 % sodium azide-7	90% H2O/10% D2O	112	6.5	ambient	293
6	3D HN(CO)CA	1 mM [U-99% 13C; U-99% 15N] entity-1, 20 mM MES-2, 100 mM sodium chloride-3, 5 mM calcium chloride-4, 10 mM DTT-5, 50 uM DSS-6, 0.02 % sodium azide-7	90% H2O/10% D2O	112	6.5	ambient	293
7	3D HNCA	1 mM [U-99% 13C; U-99% 15N] entity-1, 20 mM MES-2, 100 mM sodium chloride-3, 5 mM calcium chloride-4, 10 mM DTT-5, 50 uM DSS-6, 0.02 % sodium azide-7	90% H2O/10% D2O	112	6.5	ambient	293
8	3D H(CCO)NH	1 mM [U-99% 13C; U-99% 15N] entity-1, 20 mM MES-2, 100 mM sodium chloride-3, 5 mM calcium chloride-4, 10 mM DTT-5, 50 uM DSS-6, 0.02 % sodium azide-7	90% H2O/10% D2O	112	6.5	ambient	293
9	3D C(CO)NH	1 mM [U-99% 13C; U-99% 15N] entity-1, 20 mM MES-2, 100 mM sodium chloride-3, 5 mM calcium chloride-4, 10 mM DTT-5, 50 uM DSS-6, 0.02 % sodium azide-7	90% H2O/10% D2O	112	6.5	ambient	293
10	3D HNHA	1 mM [U-99% 13C; U-99% 15N] entity-1, 20 mM MES-2, 100 mM sodium chloride-3, 5 mM calcium chloride-4, 10 mM DTT-5, 50 uM DSS-6, 0.02 % sodium azide-7	90% H2O/10% D2O	112	6.5	ambient	293
11	3D HCCH-TOCSY	1 mM [U-99% 13C; U-99% 15N] entity-1, 20 mM MES-2, 100 mM sodium chloride-3, 5 mM calcium chloride-4, 10 mM DTT-5, 50 uM DSS-6, 0.02 % sodium azide-7	90% H2O/10% D2O	112	6.5	ambient	293
12	3D HNCO	1 mM [U-99% 13C; U-99% 15N] entity-1, 20 mM MES-2, 100 mM sodium chloride-3, 5 mM calcium chloride-4, 10 mM DTT-5, 50 uM DSS-6, 0.02 % sodium azide-7	90% H2O/10% D2O	112	6.5	ambient	293
13	3D 1H-15N NOESY	1 mM [U-99% 13C; U-99% 15N] entity-1, 20 mM MES-2, 100 mM sodium chloride-3, 5 mM calcium chloride-4, 10 mM DTT-5, 50 uM DSS-6, 0.02 % sodium azide-7	90% H2O/10% D2O	112	6.5	ambient	293
14	3D 1H-13C NOESY	1 mM [U-99% 13C; U-99% 15N] entity-1, 20 mM MES-2, 100 mM sodium chloride-3, 5 mM calcium chloride-4, 10 mM DTT-5, 50 uM DSS-6, 0.02 % sodium azide-7	90% H2O/10% D2O	112	6.5	ambient	293
15	4D-1H-13C-13C-1H HMQC-NOESY-HMQC	1 mM [U-99% 13C; U-99% 15N] entity-8, 20 mM MES-9, 100 mM sodium chloride-10, 5 mM calcium chloride-11, 10 mM DTT-12, 50 uM DSS-13, 0.02 % sodium azide-14	100% D2O	112	6.5	ambient	293

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Varian	INOVA	600
2	Bruker	DMX	850

NMR Refinement
Method	Details	Software
molecular dynamics, simulated annealing		AutoStructure

NMR Ensemble Information
Conformer Selection Criteria	lowest energy, fewest restraint violations, favorable backbone geometry
Conformers Calculated Total Number	40
Conformers Submitted Total Number	20
Representative Model	1 (combination of low energy, few restraint violations, good backbone geometry)

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	structure solution	AutoStructure		Huang, Tejero, Powers and Montelione
2	data analysis	Felix		Felix NMR Inc.
3	processing	Felix		Felix NMR Inc.
4	processing	NMRPipe		Delaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax
5	refinement	CNS		Brunger, Adams, Clore, Gros, Nilges and Read
6	refinement	X-PLOR NIH		Schwieters, Kuszewski, Tjandra and Clore
7	structure solution	PSVS		Bhattacharya and Montelione
8	data analysis	PSVS		Bhattacharya and Montelione
9	data analysis	TALOS		Cornilescu, Delaglio and Bax

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.