2MZB

Solution structural studies of GTP:adenosylcobinamide-phosphate guanylyltransferase (CobY) from Methanocaldococcus jannaschii

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	2D 1H-15N HSQC	2.0 mM [U-100% 15N] CobY, 2.0 mM [U-100% 15N] CobY, 2.0 mM [U-100% 15N] CobY, 2.0 mM [U-100% 15N] CobY, 2.0 mM [U-100% 15N] CobY, 2.0 mM [U-100% 15N] CobY	90% H2O/10% D2O	0.1	7	ambient	313
2	2D 1H-13C HSQC	2.0 mM [U-100% 13C; U-100% 15N] CobY, 2.0 mM [U-100% 13C; U-100% 15N] CobY, 2.0 mM [U-100% 13C; U-100% 15N] CobY, 2.0 mM [U-100% 13C; U-100% 15N] CobY, 2.0 mM [U-100% 13C; U-100% 15N] CobY, 2.0 mM [U-100% 13C; U-100% 15N] CobY	90% H2O/10% D2O	0.1	7	ambient	313
3	3D HNCACB	2.0 mM [U-100% 13C; U-100% 15N] CobY, 2.0 mM [U-100% 13C; U-100% 15N] CobY, 2.0 mM [U-100% 13C; U-100% 15N] CobY, 2.0 mM [U-100% 13C; U-100% 15N] CobY, 2.0 mM [U-100% 13C; U-100% 15N] CobY, 2.0 mM [U-100% 13C; U-100% 15N] CobY	90% H2O/10% D2O	0.1	7	ambient	313
4	3D CBCA(CO)NH	2.0 mM [U-100% 13C; U-100% 15N] CobY, 2.0 mM [U-100% 13C; U-100% 15N] CobY, 2.0 mM [U-100% 13C; U-100% 15N] CobY, 2.0 mM [U-100% 13C; U-100% 15N] CobY, 2.0 mM [U-100% 13C; U-100% 15N] CobY, 2.0 mM [U-100% 13C; U-100% 15N] CobY	90% H2O/10% D2O	0.1	7	ambient	313
5	3D HNCO	2.0 mM [U-100% 13C; U-100% 15N] CobY, 2.0 mM [U-100% 13C; U-100% 15N] CobY, 2.0 mM [U-100% 13C; U-100% 15N] CobY, 2.0 mM [U-100% 13C; U-100% 15N] CobY, 2.0 mM [U-100% 13C; U-100% 15N] CobY, 2.0 mM [U-100% 13C; U-100% 15N] CobY	90% H2O/10% D2O	0.1	7	ambient	313
6	3D H(CCO)NH	2.0 mM [U-100% 13C; U-100% 15N] CobY, 2.0 mM [U-100% 13C; U-100% 15N] CobY, 2.0 mM [U-100% 13C; U-100% 15N] CobY, 2.0 mM [U-100% 13C; U-100% 15N] CobY, 2.0 mM [U-100% 13C; U-100% 15N] CobY, 2.0 mM [U-100% 13C; U-100% 15N] CobY	90% H2O/10% D2O	0.1	7	ambient	313
7	3D HCCH-TOCSY	2.0 mM [U-100% 13C; U-100% 15N] CobY, 2.0 mM [U-100% 13C; U-100% 15N] CobY, 2.0 mM [U-100% 13C; U-100% 15N] CobY, 2.0 mM [U-100% 13C; U-100% 15N] CobY, 2.0 mM [U-100% 13C; U-100% 15N] CobY, 2.0 mM [U-100% 13C; U-100% 15N] CobY	90% H2O/10% D2O	0.1	7	ambient	313
8	3D 1H-15N NOESY	2.0 mM [U-100% 13C; U-100% 15N] CobY, 2.0 mM [U-100% 13C; U-100% 15N] CobY, 2.0 mM [U-100% 13C; U-100% 15N] CobY, 2.0 mM [U-100% 13C; U-100% 15N] CobY, 2.0 mM [U-100% 13C; U-100% 15N] CobY, 2.0 mM [U-100% 13C; U-100% 15N] CobY	90% H2O/10% D2O	0.1	7	ambient	313
9	3D 1H-13C NOESY aliphatic	2.0 mM [U-100% 13C; U-100% 15N] CobY, 2.0 mM [U-100% 13C; U-100% 15N] CobY, 2.0 mM [U-100% 13C; U-100% 15N] CobY, 2.0 mM [U-100% 13C; U-100% 15N] CobY, 2.0 mM [U-100% 13C; U-100% 15N] CobY, 2.0 mM [U-100% 13C; U-100% 15N] CobY	90% H2O/10% D2O	0.1	7	ambient	313

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Varian	INOVA	900
2	Varian	INOVA	800
3	Varian	INOVA	600

NMR Refinement
Method	Details	Software
DGSA-distance geometry simulated annealing, simulated annealing		X-PLOR NIH

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

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	refinement	X-PLOR NIH		Schwieters, Kuszewski, Tjandra and Clore
2	collection	VnmrJ		Varian
3	processing	NMRPipe		Delaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax
4	data analysis	XEASY		Bartels et al.
5	data analysis	TALOS		Cornilescu, Delaglio and Bax
6	data analysis	Sparky		Goddard
7	chemical shift assignment	CYANA		Guntert, Mumenthaler and Wuthrich
8	structure solution	CYANA		Guntert, Mumenthaler and Wuthrich
9	data analysis	MOLMOL		Koradi, Billeter and Wuthrich
10	structure solution	Procheck		Laskowski, MacArthur, Smith, Jones, Hutchinson, Morris, Moss and Thornton
11	refinement	CYANA		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.