Experiment: 6E4H

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	2D TROSY-HSQC	1.2 mM [U-100% 13C; U-100% 15N] PALB2cc, 20 mM MES, 200 mM sodium chloride, 5 mM Calcium Chloride, 10 mM DTT, 10 % DSS, 0.02 % sodium azide	90% H2O/10% D2O	200 mM	6.5	1 atm	298	Bruker AVANCE 800
2	2D 1H-13C HSQC aliphatic	1.2 mM [U-100% 13C; U-100% 15N] PALB2cc, 20 mM MES, 200 mM sodium chloride, 5 mM Calcium Chloride, 10 mM DTT, 10 % DSS, 0.02 % sodium azide	90% H2O/10% D2O	200 mM	6.5	1 atm	298	Bruker AVANCE 800
3	3D TROSY-HNCOCA	1.2 mM [U-100% 13C; U-100% 15N] PALB2cc, 20 mM MES, 200 mM sodium chloride, 5 mM Calcium Chloride, 10 mM DTT, 10 % DSS, 0.02 % sodium azide	90% H2O/10% D2O	200 mM	6.5	1 atm	298	Bruker AVANCE 800
4	3D TROSY-HNCA	1.2 mM [U-100% 13C; U-100% 15N] PALB2cc, 20 mM MES, 200 mM sodium chloride, 5 mM Calcium Chloride, 10 mM DTT, 10 % DSS, 0.02 % sodium azide	90% H2O/10% D2O	200 mM	6.5	1 atm	298	Bruker AVANCE 800
5	3D TROSY-HNCOCACB	1.2 mM [U-100% 13C; U-100% 15N] PALB2cc, 20 mM MES, 200 mM sodium chloride, 5 mM Calcium Chloride, 10 mM DTT, 10 % DSS, 0.02 % sodium azide	90% H2O/10% D2O	200 mM	6.5	1 atm	298	Bruker AVANCE 800
7	3D TROSY-HNCACB	1.2 mM [U-100% 13C; U-100% 15N] PALB2cc, 20 mM MES, 200 mM sodium chloride, 5 mM Calcium Chloride, 10 mM DTT, 10 % DSS, 0.02 % sodium azide	90% H2O/10% D2O	200 mM	6.5	1 atm	298	Bruker AVANCE 800
15	3D TROSY-HNCO	1.2 mM [U-100% 13C; U-100% 15N] PALB2cc, 20 mM MES, 200 mM sodium chloride, 5 mM Calcium Chloride, 10 mM DTT, 10 % DSS, 0.02 % sodium azide	90% H2O/10% D2O	200 mM	6.5	1 atm	298	Bruker AVANCE 800
6	3D TROSY-HBHACONH	1.2 mM [U-100% 13C; U-100% 15N] PALB2cc, 20 mM MES, 200 mM sodium chloride, 5 mM Calcium Chloride, 10 mM DTT, 10 % DSS, 0.02 % sodium azide	90% H2O/10% D2O	200 mM	6.5	1 atm	298	Bruker AVANCE 800
8	3D HCCH-TOCSY	1.2 mM [U-100% 13C; U-100% 15N] PALB2cc, 20 mM MES, 200 mM sodium chloride, 5 mM Calcium Chloride, 10 mM DTT, 10 % DSS, 0.02 % sodium azide	90% H2O/10% D2O	200 mM	6.5	1 atm	298	Bruker AVANCE 800
10	3D CCH-TOCSY	1.2 mM [U-100% 13C; U-100% 15N] PALB2cc, 20 mM MES, 200 mM sodium chloride, 5 mM Calcium Chloride, 10 mM DTT, 10 % DSS, 0.02 % sodium azide	90% H2O/10% D2O	200 mM	6.5	1 atm	298	Bruker AVANCE 800
9	AROMATIC CCH-TOCSY	1.2 mM [U-100% 13C; U-100% 15N] PALB2cc, 20 mM MES, 200 mM sodium chloride, 5 mM Calcium Chloride, 10 mM DTT, 10 % DSS, 0.02 % sodium azide	90% H2O/10% D2O	200 mM	6.5	1 atm	298	Bruker AVANCE 800
11	3D 1H-13C NOESY	1.2 mM [U-100% 13C; U-100% 15N] PALB2cc, 20 mM MES, 200 mM sodium chloride, 5 mM Calcium Chloride, 10 mM DTT, 10 % DSS, 0.02 % sodium azide	90% H2O/10% D2O	200 mM	6.5	1 atm	298	Bruker AVANCE 800
13	3D 1H-13C NOESY aromatic	1.2 mM [U-100% 13C; U-100% 15N] PALB2cc, 20 mM MES, 200 mM sodium chloride, 5 mM Calcium Chloride, 10 mM DTT, 10 % DSS, 0.02 % sodium azide	90% H2O/10% D2O	200 mM	6.5	1 atm	298	Bruker AVANCE 800
12	3D simultaneous 13C-aromatic,13C-aliphatic,15N edited 1H-1H NOESY	1.2 mM [U-100% 13C; U-100% 15N] PALB2cc, 20 mM MES, 200 mM sodium chloride, 5 mM Calcium Chloride, 10 mM DTT, 10 % DSS, 0.02 % sodium azide	90% H2O/10% D2O	200 mM	6.5	1 atm	298	Bruker AVANCE 800
14	X-filtered noesy (pulse sequence: noesyhsqcgpwgx13d)	1 v/v [U-100% 13C; U-100% 15N] PALB2cc, 2 v/v unlabeled PALB2cc, 20 mM MES, 200 mM sodium chloride, 5 mM Calcium Chloride, 10 mM DTT, 0.02 % sodium azide, 10 % DSS	90% H2O/10% D2O	200 mM	6.5	1 atm	298	Bruker AVANCE 800

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

NMR Refinement
Method	Details	Software
simulated annealing	molecular dynamics	CNS

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

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	processing	NMRPipe		Delaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax
2	processing	NMRDraw		Delaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax
3	collection	TopSpin		Bruker Biospin
4	peak picking	Sparky		Goddard
5	chemical shift assignment	AutoAssign		Zimmerman, Moseley, Kulikowski and Montelione
6	chemical shift assignment	PINE		Bahrami, Markley, Assadi, and Eghbalnia
7	structure calculation	CYANA		Guntert, Mumenthaler and Wuthrich
8	refinement	CNS		Brunger, Adams, Clore, Gros, Nilges and Read
9	chemical shift assignment	AVS		Moseley and Montelione
10	data analysis	TALOS		Cornilescu, Delaglio and Bax
11	data analysis	PSVS		Bhattacharya and Montelione
12	chemical shift assignment	XEASY		Bartels et al.

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.