Experiment: 2L4H

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	2D 1H-15N HSQC IPAP	0.5 mM [U-13C; U-15N; U-2H] CIB1, 50 mM HEPES, 100 mM potassium chloride, 4 mM calcium cloride, 10 mM DTT	90% H2O/10% D2O	200	7.5	ambient	310
2	3D HNCO type for CaCO and C'N RDC	0.5 mM [U-13C; U-15N; U-2H] CIB1, 50 mM HEPES, 100 mM potassium chloride, 4 mM calcium cloride, 10 mM DTT	90% H2O/10% D2O	200	7.5	ambient	310
3	3D 1H-15N NOESY	0.5 mM I/L/V methyl 13C and 1H, [U,2H] CIB1, 50 mM HEPES, 100 mM potassium chloride, 4 mM calcium chloride, 10 mM DTT	90% H2O/10% D2O	200	7.5	ambient	310
4	3D 1H-13C NOESY	0.5 mM I/L/V methyl 13C and 1H, [U,2H] CIB1, 50 mM HEPES, 100 mM potassium chloride, 4 mM calcium chloride, 10 mM DTT	90% H2O/10% D2O	200	7.5	ambient	310

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Bruker	AVANCE	700

NMR Refinement
Method	Details	Software
simulated annealing	two steps: 1st step, 200K-20K; 2nd step, 20K-2K	X-PLOR NIH

NMR Ensemble Information
Conformer Selection Criteria	structures with the lowest energy
Conformers Calculated Total Number	100
Conformers Submitted Total Number	11
Representative Model	1 (lowest energy)

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	refinement	X-PLOR NIH	2.18	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.