Experiment: 2MHH

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	2D 1H-15N HSQC	0.7 mM [U-100% 13C; U-100% 15N] protein, 25 mM TRIS, 150 mM sodium chloride, 20 mM calcium chloride, 5 mM TCEP, .05 % sodium azide	95% H2O/5% D2O	0.195	7.38	ambient	298
2	2D 1H-13C HSQC	0.7 mM [U-100% 13C; U-100% 15N] protein, 25 mM TRIS, 150 mM sodium chloride, 20 mM calcium chloride, 5 mM TCEP, .05 % sodium azide	95% H2O/5% D2O	0.195	7.38	ambient	298
3	3D HNCO	0.7 mM [U-100% 13C; U-100% 15N] protein, 25 mM TRIS, 150 mM sodium chloride, 20 mM calcium chloride, 5 mM TCEP, .05 % sodium azide	95% H2O/5% D2O	0.195	7.38	ambient	298
4	3D HNCACB	0.7 mM [U-100% 13C; U-100% 15N] protein, 25 mM TRIS, 150 mM sodium chloride, 20 mM calcium chloride, 5 mM TCEP, .05 % sodium azide	95% H2O/5% D2O	0.195	7.38	ambient	298
5	3D HN(CO)CA	0.7 mM [U-100% 13C; U-100% 15N] protein, 25 mM TRIS, 150 mM sodium chloride, 20 mM calcium chloride, 5 mM TCEP, .05 % sodium azide	95% H2O/5% D2O	0.195	7.38	ambient	298
6	3D HN(CA)CO	0.7 mM [U-100% 13C; U-100% 15N] protein, 25 mM TRIS, 150 mM sodium chloride, 20 mM calcium chloride, 5 mM TCEP, .05 % sodium azide	95% H2O/5% D2O	0.195	7.38	ambient	298
7	3D HCACO	0.7 mM [U-100% 13C; U-100% 15N] protein, 25 mM TRIS, 150 mM sodium chloride, 20 mM calcium chloride, 5 mM TCEP, .05 % sodium azide	95% H2O/5% D2O	0.195	7.38	ambient	298
8	3D C(CO)NH	0.7 mM [U-100% 13C; U-100% 15N] protein, 25 mM TRIS, 150 mM sodium chloride, 20 mM calcium chloride, 5 mM TCEP, .05 % sodium azide	95% H2O/5% D2O	0.195	7.38	ambient	298
9	3D 1H-15N TOCSY	0.7 mM [U-100% 13C; U-100% 15N] protein, 25 mM TRIS, 150 mM sodium chloride, 20 mM calcium chloride, 5 mM TCEP, .05 % sodium azide	95% H2O/5% D2O	0.195	7.38	ambient	298
10	3D HCCH-TOCSY	0.7 mM [U-100% 13C; U-100% 15N] protein, 25 mM TRIS, 150 mM sodium chloride, 20 mM calcium chloride, 5 mM TCEP, .05 % sodium azide	95% H2O/5% D2O	0.195	7.38	ambient	298
11	3D 1H-15N NOESY	0.7 mM [U-100% 13C; U-100% 15N] protein, 25 mM TRIS, 150 mM sodium chloride, 20 mM calcium chloride, 5 mM TCEP, .05 % sodium azide	95% H2O/5% D2O	0.195	7.38	ambient	298
12	3D 1H-13C NOESY aliphatic	0.7 mM [U-100% 13C; U-100% 15N] protein, 25 mM TRIS, 150 mM sodium chloride, 20 mM calcium chloride, 5 mM TCEP, .05 % sodium azide	95% H2O/5% D2O	0.195	7.38	ambient	298
13	3D 1H-13C NOESY aromatic	0.7 mM [U-100% 13C; U-100% 15N] protein, 25 mM TRIS, 150 mM sodium chloride, 20 mM calcium chloride, 5 mM TCEP, .05 % sodium azide	95% H2O/5% D2O	0.195	7.38	ambient	298
14	2D 1H-15N HSQC	0.7 mM [U-100% 13C; U-100% 15N] protein, 25 mM TRIS, 150 mM sodium chloride, 20 mM calcium chloride, 5 mM TCEP, .05 % sodium azide	95% H2O/5% D2O	0.195	7.38	ambient	298

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Varian	INOVA	600

NMR Refinement
Method	Details	Software
simulated annealing		VNMR

NMR Ensemble Information
Conformer Selection Criteria	structures with the lowest energy
Conformers Calculated Total Number	80
Conformers Submitted Total Number	20
Representative Model	1 (lowest energy)

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	collection	VNMR		Varian
2	chemical shift assignment	Sparky		Goddard
3	peak picking	Sparky		Goddard
4	processing	NMRPipe		Delaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax
5	data analysis	TALOS		Cornilescu, Delaglio and Bax
6	data analysis	CYANA		Guntert, Mumenthaler and Wuthrich
7	refinement	X-PLOR NIH		Schwieters, Kuszewski, Tjandra and Clore

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.