2EZN
SOLUTION NMR STRUCTURE OF CYANOVIRIN-N ENSEMBLE OF 40 SIMULATED ANNEALING STRUCTURES
SOLUTION NMR
| NMR Experiment | ||||||||
|---|---|---|---|---|---|---|---|---|
| Experiment | Type | Sample Contents | Solvent | Ionic Strength | pH | Pressure | Temperature (K) | Spectrometer |
| 1 | TRIPLE RESONANCE FOR ASSIGNMENT OF PROTEIN: CBCA(CO)NH | 6.1 | 300 | |||||
| 2 | CBCANH | 6.1 | 300 | |||||
| 3 | HNCO | 6.1 | 300 | |||||
| 4 | C(CO)NH | 6.1 | 300 | |||||
| 5 | H(CCO)NH | 6.1 | 300 | |||||
| 6 | HCCH-COSY | 6.1 | 300 | |||||
| 7 | HCCH-TOCSY | 6.1 | 300 | |||||
| 8 | HNHA | 6.1 | 300 | |||||
| 9 | 15N-SEPARATED HOHAHA; QUANTITATIVE J CORRELATION FOR COUPLING CONSTANTS; 3D 15N-SEPARATED NOE | 6.1 | 300 | |||||
| 10 | 3D 13C-SEPARATED NOE AND ROE | 6.1 | 300 | |||||
| 11 | 4D 15N/13C-SEPARATED NOE | 6.1 | 300 | |||||
| 12 | 4D 13C/13C-SEPARATED NOE EXPERIMENTS; 3D HCA(CO)N FOR THREE-BOND AMINIDE DEUTERIUM ISOTOPE SHIFTS; VARIOUS COUPLED 2D AND 3D SPECTRA TO MEASURE THE N-H | 6.1 | 300 | |||||
| 13 | CA-H | 6.1 | 300 | |||||
| 14 | C-H CA-C' | 6.1 | 300 | |||||
| 15 | N-C' | 6.1 | 300 | |||||
| 16 | HN-C' DIPOLAR COUPLINGS OBTAINED BY TAKING THE DIFFERENCE IN THE J SPLITTINGS IN ISOTROPIC MEDIUM | 6.1 | 300 | |||||
| 17 | IN A LIQUID CRYSTALLINE MEDIUM (4% 3:1 DMPC:DHPC). | 6.1 | 300 | |||||
| NMR Spectrometer Information | |||
|---|---|---|---|
| Spectrometer | Manufacturer | Model | Field Strength |
| 1 | Bruker | DMX500 | 500 |
| 2 | Bruker | DMX600 | 600 |
| 3 | Bruker | DMX750 | 750 |
| NMR Refinement | ||
|---|---|---|
| Method | Details | Software |
| simulated annealing | AVE.RMS DIFF. TO MEAN FOR ALL NON-H-ATOMS (RESIDUES 1:101)= 0.433636 AVE.RMS DIFF. TO MEAN FOR BACKBONE ATOMS (N, CA, C', O) (RESIDUES 1:101)= 0.139826 RMS DEVIATIONS FOR BONDS, ANGLES, IMPROPERS, CDIH, NOE, COUP 5.067337E-03, 0.712983, 0.667194, 0.157308, 1.428009E-02, 0.608909 C13CA AND CB SHIFTS RMS : 0.852581, 1.15742 JCOUP STATS: NON-GLY RESIDUES GLY RMS-D: 0.608909 1.46813 BACKBONE DIPOLAR COUPLINGS NH CH CACO NCO HNCO RMS : 0.466712 1.15058 1.29414 0.572019 1.2653 SIDECHAIN DIPOLAR COUPLINGS CH CH3S CH3D ARO RMS DIPO_SIDE: 1.6875 0.796796 0.531907 0.160016 RMS FOR 1H SHIFTS: ALL ALPHA ALPHA_GLY METHYL(S) METHYL(D) OTHER(S) OTHER(D) RMS PROT: 0.263524 0.243015 0.23853 0.115892 0.148148 0.267304 0.300122 IN THE RESTRAINED REGULARIZED MEAN COORDINATES (2EZM) THE LAST COLUMN REPRESENTS THE AVERAGE RMS DIFFERENCE BETWEEN THE INDIVIDUAL SIMULATED ANNEALING STRUCTURES AND THE MEAN COORDINATE POSITIONS. THE LAST COLUMN IN THE INDIVIDUAL SA STRUCTURES (2EZN) HAS NO MEANING. BEST FITTING TO GENERATE THE AVERAGE STRUCTURE IS WITH RESPECT TO RESIDUES 1-101. NOTE THE OCCUPANCY FIELD HAS NO MEANING. | CNS |
| NMR Ensemble Information | |
|---|---|
| Conformer Selection Criteria | |
| Conformers Calculated Total Number | |
| Conformers Submitted Total Number | 40 |
| Additional NMR Experimental Information | |
|---|---|
| Details | THE 3D STRUCTURE OF CYANOVIRIN SOLVED BY MULTI-DIMENSIONAL HETERONUCLEAR NMR AND IS BASED ON 2597 EXPERIMENTAL NMR RESTRAINTS: 419 SEQUENTIAL (|I- J|=1), 170 MEDIUM RANGE (1 < |I-J| <=5) AND 554 LONG RANGE (|I-J| >5) INTERRESIDUES AND 19 INTRARESIDUE APPROXIMATE INTERPROTON DISTANCE RESTRAINTS; 109 DISTANCE RESTRAINTS FOR 55 H-BONDS; 339 TORSION ANGLE RESTRAINTS (100 PHI, 98 PSI, 76 CHI1, 48 CHI2, 15 CHI3, 2 CHI4); 82 THREE-BOND HN-HA COUPLING CONSTANT RESTRAINTS; 157 (82 CALPHA AND 75 CBETA) 13C SHIFT RESTRAINTS; 362 1H SHIFT RESTRAINTS; AND 386 DIPOLAR COUPLING RESTRAINTS (82 N-H, 76 C-H, 43 CA-C', 65 N-C' 62 HNC', 58 SIDE-CHAIN C-H). THE STRUCTURES WERE CALCULATED USING THE SIMULATED ANNEALING PROTOCOL OF NILGES ET AL. (1988) FEBS LETT. 229, 129-136 USING THE PROGRAM CNS (BRUNGER ET AL. ACTA CRYST SERIES D IN PRESS) MODIFIED TO INCORPORATE COUPLING CONSTANT (GARRETT ET AL. (1984) J. MAGN. RESON. SERIES B 104, 99-103), CARBON CHEMICAL SHIFT (KUSZEWSKI ET AL. (1995) J. MAGN. RESON. SERIES B 106, 92-96), 1H CHEMICAL SHIFT (KUSZEWSKI ET AL. (1995) J. MAGN. RESON. SERIES B 107, 293-297; KUSZEWSKI ET AL. (1996) J. MAGN. RESON. SERIES B 112, 79-81), AND DIPOLAR COUPLING (CLORE ET AL. (1998) J. MAGN. RESON. 131, 159-162) RESTRAINTS, AND A CONFORMATIONAL DATABASE POTENTIAL (KUSZEWSKI ET AL. (1996) PROTEIN SCI. 5, 1067-1080; KUSZEWSKI ET AL. (1997) J. MAGN. RESON 125, 171-177). |
| Computation: NMR Software | ||||
|---|---|---|---|---|
| # | Classification | Version | Software Name | Author |
| 1 | refinement | CNS | BRUNGER, ADAMS, CLORE, DELANO, GROSSE-KUNSTLEVE,JIANG,KUSZEWSKI,NILGES, PANNU,READ,RICE,SIMONSON,WARREN | |
| 2 | structure solution | CNS (SEE ABOVE) | ABOVE) | |
