1EQK

SOLUTION STRUCTURE OF ORYZACYSTATIN-I, A CYSTEINE PROTEINASE INHIBITOR OF THE RICE, ORYZA SATIVA L. JAPONICA

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	2D NOESY	3 mM oryzacystatin-I; 50 mM sodium phosphate/100 mM NaCl/0.02% NaN3, pH 6.8; 10% D2O/90% H2O	10% D2O/90% H2O		6.8	ambient	298
2	3D_15N-separated_NOESY	2 mM oryzacystatin-I U-15N; 50 mM sodium phosphate/100 mM NaCl/0.02% NaN3, pH 6.8; 10% D2O/90% H2O	10% D2O/90% H2O		6.8	ambient	298
3	HNHA	2 mM oryzacystatin-I U-15N; 50 mM sodium phosphate/100 mM NaCl/0.02% NaN3, pH 6.8; 10% D2O/90% H2O	10% D2O/90% H2O		6.8	ambient	298

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Varian	INOVA	500

NMR Refinement
Method	Details	Software
torsion angle dynamics	A total of 1383 restraints, consisting of 1183 interproton distance restraints (338 long-range (\|i - j\| >= 6), 228 medium-range (2 <= \|i - j\| <= 5), 351 sequential (\|i - j\| = 1) and 266 intraresidual (\|i - j\| = 0)), 108 hydrogen bond restraints (representing 54 hydrogen bonds) and 92 torsion angle restraints (54 phi and 38 chi1), were used in the structure calculations by torsion angle dynamics using DYANA (ver. 1.4). A final set of 20 structures was selected from 100 calculations on the basis of agreement with the experimental data and van der Waals' energy. The average coordinates of the 20 DYANA structures were subjected to energy-minimization using CNS (ver. 0.9).	VNMR

NMR Ensemble Information
Conformer Selection Criteria	structures with favorable non-bond energy
Conformers Calculated Total Number	100
Conformers Submitted Total Number	21
Representative Model	21 (minimized average structure)

Additional NMR Experimental Information
Details	The structure was determined using 1H and 15N NMR spectroscopy.

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	collection	VNMR	5.3B	Varian
2	processing	NMRPipe	1.6	Delaglio, F. et al.
3	data analysis	NMRDraw	1.6	Delaglio, F. et al.
4	data analysis	Felix	95	Molecular Simulations
5	data analysis	PIPP/CAPP	4.0	Garrett, D. et al.
6	structure solution	DYANA	1.4	Guentert, P. et al.
7	refinement	CNS	0.9	Brunger,ADAMS,CLORE,DELANO,GROS,GROSSE-KUNSTLEVE,JIANG,KUSZEWSKI,NILGES,PANNU,READ,RICE,SIMONSON,WARREN
8	data analysis	AQUA	2.0	Rullmann, J.A.C. et al.
9	data analysis	PROCHECK-NMR	3.4	Laskowski, R.A. et al.
10	data analysis	MOLMOL	2.6	Koradi, R. 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.