Experiment: 2L6A

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	2D 1H-15N HSQC	600 uM [U-99% 15N] NLRP12 PYD-1, 20 mM sodium phosphate-2, 500 mM sodium chloride-3, 0.5 mM TCEP-4	90% H2O/10% D2O	0.5	6.5	ambient	298
2	2D 1H-13C HSQC	600 uM [U-99% 13C; U-99% 15N] NLRP12 PYD-5, 20 mM sodium phosphate-6, 500 mM sodium chloride-7, 0.5 mM TCEP-8	90% H2O/10% D2O	0.5	6.5	ambient	298
3	3D HNCA	600 uM [U-99% 13C; U-99% 15N] NLRP12 PYD-5, 20 mM sodium phosphate-6, 500 mM sodium chloride-7, 0.5 mM TCEP-8	90% H2O/10% D2O	0.5	6.5	ambient	298
4	3D HNCACB	600 uM [U-99% 13C; U-99% 15N] NLRP12 PYD-5, 20 mM sodium phosphate-6, 500 mM sodium chloride-7, 0.5 mM TCEP-8	90% H2O/10% D2O	0.5	6.5	ambient	298
5	3D CBCA(CO)NH	600 uM [U-99% 13C; U-99% 15N] NLRP12 PYD-5, 20 mM sodium phosphate-6, 500 mM sodium chloride-7, 0.5 mM TCEP-8	90% H2O/10% D2O	0.5	6.5	ambient	298
6	3D HNCO	600 uM [U-99% 13C; U-99% 15N] NLRP12 PYD-5, 20 mM sodium phosphate-6, 500 mM sodium chloride-7, 0.5 mM TCEP-8	90% H2O/10% D2O	0.5	6.5	ambient	298
7	3D HN(CA)CO	600 uM [U-99% 13C; U-99% 15N] NLRP12 PYD-5, 20 mM sodium phosphate-6, 500 mM sodium chloride-7, 0.5 mM TCEP-8	90% H2O/10% D2O	0.5	6.5	ambient	298
8	3D HBHA(CO)NH	600 uM [U-99% 13C; U-99% 15N] NLRP12 PYD-5, 20 mM sodium phosphate-6, 500 mM sodium chloride-7, 0.5 mM TCEP-8	90% H2O/10% D2O	0.5	6.5	ambient	298
9	3D C(CO)NH	600 uM [U-99% 13C; U-99% 15N] NLRP12 PYD-5, 20 mM sodium phosphate-6, 500 mM sodium chloride-7, 0.5 mM TCEP-8	90% H2O/10% D2O	0.5	6.5	ambient	298
10	3D HCCH-TOCSY	600 uM [U-99% 13C; U-99% 15N] NLRP12 PYD-5, 20 mM sodium phosphate-6, 500 mM sodium chloride-7, 0.5 mM TCEP-8	90% H2O/10% D2O	0.5	6.5	ambient	298
11	3D 1H-15N NOESY	600 uM [U-99% 15N] NLRP12 PYD-1, 20 mM sodium phosphate-2, 500 mM sodium chloride-3, 0.5 mM TCEP-4	90% H2O/10% D2O	0.5	6.5	ambient	298
12	3D 1H-13C NOESY	600 uM [U-99% 13C; U-99% 15N] NLRP12 PYD-5, 20 mM sodium phosphate-6, 500 mM sodium chloride-7, 0.5 mM TCEP-8	90% H2O/10% D2O	0.5	6.5	ambient	298
13	2D 1H-1H NOESY	600 uM NLRP12 PYD-9, 20 mM sodium phosphate-10, 500 mM sodium chloride-11, 0.5 mM TCEP-12	100% D2O	0.5	6.5	ambient	298
14	2D 1H-1H TOCSY	600 uM NLRP12 PYD-9, 20 mM sodium phosphate-10, 500 mM sodium chloride-11, 0.5 mM TCEP-12	100% D2O	0.5	6.5	ambient	298
15	2D 1H-1H COSY	600 uM NLRP12 PYD-9, 20 mM sodium phosphate-10, 500 mM sodium chloride-11, 0.5 mM TCEP-12	100% D2O	0.5	6.5	ambient	298

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Bruker	AVANCE	500
2	Bruker	AVANCE	800

NMR Refinement
Method	Details	Software
simulated annealing	Refinement was done in explicit solvent using CNS and the RECOORD script package.	TopSpin

NMR Ensemble Information
Conformer Selection Criteria	structures with the lowest energy
Conformers Calculated Total Number	100
Conformers Submitted Total Number	20
Representative Model	1 (lowest energy)

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	collection	TopSpin	2.1	Bruker Biospin
2	processing	TopSpin	2.1	Bruker Biospin
3	chemical shift assignment	CARA		Keller and Wuthrich
4	structure solution	CYANA	2.1	Guntert, Mumenthaler and Wuthrich
5	refinement	CNS		Brunger, Adams, Clore, Gros, Nilges and Read

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.