Experiment: 6DRF

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	2D 1H-15N HSQC	0.9 mM [U-15N] human RD3, 20 mM D11 TRIS, 3 mM D10 DTT, 50 uM D12 EDTA, 0.04 % w/v sodium azide	93% H2O/7% D2O	20 mM	7.4	1 atm	296	Bruker AVANCE 600
2	3D HNCA	0.9 mM [U-13C; U-15N] human RD3, 20 mM d11 TRIS, 3 mM d10 DTT, 50 uM d12 EDTA, 0.04 % w/v sodium azide	93% H2O/7% D2O	20 mM	7.4	1 atm	296	Bruker AVANCE 600
3	3D HNCACB	0.9 mM [U-13C; U-15N] human RD3, 20 mM d11 TRIS, 3 mM d10 DTT, 50 uM d12 EDTA, 0.04 % w/v sodium azide	93% H2O/7% D2O	20 mM	7.4	1 atm	296	Bruker AVANCE 600
4	3D CBCA(CO)NH	0.9 mM [U-13C; U-15N] human RD3, 20 mM d11 TRIS, 3 mM d10 DTT, 50 uM d12 EDTA, 0.04 % w/v sodium azide	93% H2O/7% D2O	20 mM	7.4	1 atm	296	Bruker AVANCE 600
5	3D HNCO	0.9 mM [U-13C; U-15N] human RD3, 20 mM d11 TRIS, 3 mM d10 DTT, 50 uM d12 EDTA, 0.04 % w/v sodium azide	93% H2O/7% D2O	20 mM	7.4	1 atm	296	Bruker AVANCE 600
7	3D HCCH-TOCSY	0.9 mM [U-13C; U-15N] human RD3, 20 mM d11 TRIS, 3 mM d10 DTT, 50 uM d12 EDTA, 0.04 % w/v sodium azide	93% H2O/7% D2O	20 mM	7.4	1 atm	296	Bruker AVANCE 600
6	3D CCCONH	0.9 mM [U-13C; U-15N] human RD3, 20 mM d11 TRIS, 3 mM d10 DTT, 50 uM d12 EDTA, 0.04 % w/v sodium azide	93% H2O/7% D2O	20 mM	7.4	1 atm	296	Bruker AVANCE 600
8	3D HCCCONH	0.9 mM [U-13C; U-15N] human RD3, 20 mM d11 TRIS, 3 mM d10 DTT, 50 uM d12 EDTA, 0.04 % w/v sodium azide	93% H2O/7% D2O	20 mM	7.4	1 atm	296	Bruker AVANCE 600
9	3D 1H-15N NOESY	0.9 mM [U-15N] human RD3, 20 mM D11 TRIS, 3 mM D10 DTT, 50 uM D12 EDTA, 0.04 % w/v sodium azide	93% H2O/7% D2O	20 mM	7.4	1 atm	296	Bruker AVANCE 600
10	3D 1H-13C NOESY	0.9 mM [U-13C; U-15N] human RD3, 20 mM d11 TRIS, 3 mM d10 DTT, 50 uM d12 EDTA, 0.04 % w/v sodium azide	93% H2O/7% D2O	20 mM	7.4	1 atm	296	Bruker AVANCE 600
11	3D 1H-15N TOCSY	0.9 mM [U-15N] human RD3, 20 mM D11 TRIS, 3 mM D10 DTT, 50 uM D12 EDTA, 0.04 % w/v sodium azide	93% H2O/7% D2O	20 mM	7.4	1 atm	296	Bruker AVANCE 600
12	2D 1H-13C HSQC	0.9 mM [U-13C; U-15N] human RD3, 20 mM d11 TRIS, 3 mM d10 DTT, 50 uM d12 EDTA, 0.04 % w/v sodium azide	93% H2O/7% D2O	20 mM	7.4	1 atm	296	Bruker AVANCE 600

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Bruker	AVANCE	600

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

NMR Ensemble Information
Conformer Selection Criteria	structures with the lowest energy
Conformers Calculated Total Number	200
Conformers Submitted Total Number	10
Representative Model	1 (lowest energy)

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	processing	NMRPipe	8.9	Delaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax
2	chemical shift assignment	Sparky	3.134	Goddard
3	refinement	X-PLOR NIH	2.47	Schwieters, Kuszewski, Tjandra and Clore
4	structure calculation	X-PLOR NIH	2.47	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.