1LMJ

NMR Study of the Fibrillin-1 cbEGF12-13 Pair of Ca2+ Binding Epidermal Growth Factor-like Domains

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	2D NOESY	20 mM CaCl2, 4.55 mM Tris, 3.6 mM protein	99.9% D2O	20 mM CaCl2, 4.55 mM Tris	6.5	ambient	306
2	3D_15N-separated_NOESY	20 mM CaCl2, 4.55 mM Tris, 3.8 mM 15N-protein	90% H2O/10% D2O	20 mM CaCl2, 4.55 mM Tris	6.5	ambient	306
3	HMQC-J	20 mM CaCl2, 4.55 mM Tris, 3.8 mM 15N-protein	90% H2O/10% D2O	20 mM CaCl2, 4.55 mM Tris	6.5	ambient	306
4	HSQC (slow HN)	20 mM CaCl2, 4.55 mM Tris, 3.8 mM 15N-protein	90% H2O/10% D2O	20 mM CaCl2, 4.55 mM Tris	6.5	ambient	306

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	GE	OMEGA	500
2	GE	OMEGA	600
3	GE	OMEGA	750

NMR Refinement
Method	Details	Software
simulated annealing	1892 distance constraints including 411 ambiguous constraints, 26 torsion angle phi restraints, 24 restraints for 12 hydrogen bonds RMSD from experimental restraints All 0.013+/-0.001 (1932) Intraresidue 0.009+/-0.002 (504) Sequential 0.011+/-0.002 (388) Short-range (i-j<=4) (211) 0.015+/-0.003 Long-range 0.013+/-0.002 (378) Ambiguous 0.015+/-0.002 (411) H-bonds 0.014+/-0.004 (24) Calcium 0.012+/-0.005 (16) RMSD Phi rest. 0.177+/-0.093 (26)	X-PLOR

NMR Ensemble Information
Conformer Selection Criteria	structures with acceptable covalent geometry
Conformers Calculated Total Number	100
Conformers Submitted Total Number	25
Representative Model	6 (closest to the average)

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	structure solution	X-PLOR	3.81	Brunger
2	refinement	X-PLOR	3.81	Brunger

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.