Experiment: 5ZMR

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	2D 1H-15N HSQC	0.3 mM [U-95% 13C; U-95% 15N] 26S proteasome regulatory subunit Rpn5, 10 % v/v [U-99% 2H] D2O, 0.1 mg/mL DSS, 1 mM DTT, 50 mM sodium phosphate, 50 mM sodium sulfate, 2 % v/v trifluoroethanol, 5 % w/v sucrose	90% H2O/10% D2O	50 mM sodium phosphate buffer, 50 mM Na2SO4 mM	6.0	ambient Pa	298	Bruker AVANCE III 500
2	2D 1H-13C HSQC	0.3 mM [U-95% 13C; U-95% 15N] 26S proteasome regulatory subunit Rpn5, 10 % v/v [U-99% 2H] D2O, 0.1 mg/mL DSS, 1 mM DTT, 50 mM sodium phosphate, 50 mM sodium sulfate, 2 % v/v trifluoroethanol, 5 % w/v sucrose	90% H2O/10% D2O	50 mM sodium phosphate buffer, 50 mM Na2SO4 mM	6.0	ambient Pa	298	Bruker AVANCE III 500
3	3D HNCA	0.3 mM [U-95% 13C; U-95% 15N] 26S proteasome regulatory subunit Rpn5, 10 % v/v [U-99% 2H] D2O, 0.1 mg/mL DSS, 1 mM DTT, 50 mM sodium phosphate, 50 mM sodium sulfate, 2 % v/v trifluoroethanol, 5 % w/v sucrose	90% H2O/10% D2O	50 mM sodium phosphate buffer, 50 mM Na2SO4 mM	6.0	ambient Pa	298	Bruker AVANCE III 500
4	3D HN(CO)CA	0.3 mM [U-95% 13C; U-95% 15N] 26S proteasome regulatory subunit Rpn5, 10 % v/v [U-99% 2H] D2O, 0.1 mg/mL DSS, 1 mM DTT, 50 mM sodium phosphate, 50 mM sodium sulfate, 2 % v/v trifluoroethanol, 5 % w/v sucrose	90% H2O/10% D2O	50 mM sodium phosphate buffer, 50 mM Na2SO4 mM	6.0	ambient Pa	298	Bruker AVANCE III 500
5	3D HNCO	0.3 mM [U-95% 13C; U-95% 15N] 26S proteasome regulatory subunit Rpn5, 10 % v/v [U-99% 2H] D2O, 0.1 mg/mL DSS, 1 mM DTT, 50 mM sodium phosphate, 50 mM sodium sulfate, 2 % v/v trifluoroethanol, 5 % w/v sucrose	90% H2O/10% D2O	50 mM sodium phosphate buffer, 50 mM Na2SO4 mM	6.0	ambient Pa	298	Bruker AVANCE III 500
7	3D HN(CA)CO	0.3 mM [U-95% 13C; U-95% 15N] 26S proteasome regulatory subunit Rpn5, 10 % v/v [U-99% 2H] D2O, 0.1 mg/mL DSS, 1 mM DTT, 50 mM sodium phosphate, 50 mM sodium sulfate, 2 % v/v trifluoroethanol, 5 % w/v sucrose	90% H2O/10% D2O	50 mM sodium phosphate buffer, 50 mM Na2SO4 mM	6.0	ambient Pa	298	Bruker AVANCE III 500
6	3D HNCACB	0.3 mM [U-95% 13C; U-95% 15N] 26S proteasome regulatory subunit Rpn5, 10 % v/v [U-99% 2H] D2O, 0.1 mg/mL DSS, 1 mM DTT, 50 mM sodium phosphate, 50 mM sodium sulfate, 2 % v/v trifluoroethanol, 5 % w/v sucrose	90% H2O/10% D2O	50 mM sodium phosphate buffer, 50 mM Na2SO4 mM	6.0	ambient Pa	298	Bruker AVANCE III 500
8	3D CBCA(CO)NH	0.3 mM [U-95% 13C; U-95% 15N] 26S proteasome regulatory subunit Rpn5, 10 % v/v [U-99% 2H] D2O, 0.1 mg/mL DSS, 1 mM DTT, 50 mM sodium phosphate, 50 mM sodium sulfate, 2 % v/v trifluoroethanol, 5 % w/v sucrose	90% H2O/10% D2O	50 mM sodium phosphate buffer, 50 mM Na2SO4 mM	6.0	ambient Pa	298	Bruker AVANCE III 500
10	3D HBHA(CO)NH	0.3 mM [U-95% 13C; U-95% 15N] 26S proteasome regulatory subunit Rpn5, 10 % v/v [U-99% 2H] D2O, 0.1 mg/mL DSS, 1 mM DTT, 50 mM sodium phosphate, 50 mM sodium sulfate, 2 % v/v trifluoroethanol, 5 % w/v sucrose	90% H2O/10% D2O	50 mM sodium phosphate buffer, 50 mM Na2SO4 mM	6.0	ambient Pa	298	Bruker AVANCE III 500
9	3D HCCH-COSY	0.3 mM [U-95% 13C; U-95% 15N] 26S proteasome regulatory subunit Rpn5, 10 % v/v [U-99% 2H] D2O, 0.1 mg/mL DSS, 1 mM DTT, 50 mM sodium phosphate, 50 mM sodium sulfate, 2 % v/v trifluoroethanol, 5 % w/v sucrose	90% H2O/10% D2O	50 mM sodium phosphate buffer, 50 mM Na2SO4 mM	6.0	ambient Pa	298	Bruker AVANCE III 500
11	3D HCCH-COSY	0.3 mM [U-95% 13C; U-95% 15N] 26S proteasome regulatory subunit Rpn5, 10 % v/v [U-99% 2H] D2O, 0.1 mg/mL DSS, 1 mM DTT, 50 mM sodium phosphate, 50 mM sodium sulfate, 2 % v/v trifluoroethanol, 5 % w/v sucrose	90% H2O/10% D2O	50 mM sodium phosphate buffer, 50 mM Na2SO4 mM	6.0	ambient Pa	298	Bruker AVANCE III 500
12	3D 15N-edited TOCSY-HSQC	0.3 mM [U-95% 13C; U-95% 15N] 26S proteasome regulatory subunit Rpn5, 10 % v/v [U-99% 2H] D2O, 0.1 mg/mL DSS, 1 mM DTT, 50 mM sodium phosphate, 50 mM sodium sulfate, 2 % v/v trifluoroethanol, 5 % w/v sucrose	90% H2O/10% D2O	50 mM sodium phosphate buffer, 50 mM Na2SO4 mM	6.0	ambient Pa	298	Bruker AVANCE III 500
13	3D 1H-15N NOESY	0.3 mM [U-95% 13C; U-95% 15N] 26S proteasome regulatory subunit Rpn5, 10 % v/v [U-99% 2H] D2O, 0.1 mg/mL DSS, 1 mM DTT, 50 mM sodium phosphate, 50 mM sodium sulfate, 2 % v/v trifluoroethanol, 5 % w/v sucrose	90% H2O/10% D2O	50 mM sodium phosphate buffer, 50 mM Na2SO4 mM	6.0	ambient Pa	298	Bruker AVANCE III 950
14	3D 1H-15C NOESY	0.3 mM [U-95% 13C; U-95% 15N] 26S proteasome regulatory subunit Rpn5, 10 % v/v [U-99% 2H] D2O, 0.1 mg/mL DSS, 1 mM DTT, 50 mM sodium phosphate, 50 mM sodium sulfate, 2 % v/v trifluoroethanol, 5 % w/v sucrose	90% H2O/10% D2O	50 mM sodium phosphate buffer, 50 mM Na2SO4 mM	6.0	ambient Pa	298	Bruker AVANCE III 950

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Bruker	AVANCE III	500
2	Bruker	AVANCE III	950

NMR Refinement
Method	Details	Software
simulated annealing		Amber

NMR Ensemble Information
Conformer Selection Criteria	20 structures for 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	refinement	Amber	14	Case, Darden, Cheatham III, Simmerling, Wang, Duke, Luo, ... and Kollman
2	structure calculation	CYANA		Guntert, Mumenthaler and Wuthrich
3	chemical shift assignment	NMRView	5.0.4	Johnson, One Moon Scientific
4	peak picking	NMRView	5.0.4	Johnson, One Moon Scientific
5	processing	NMRPipe		Delaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax

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.