THE SOLUTION STRUCTURE OF THE HEME BINDING VARIANT ARG98CYS OF OXIDIZED ESCHERICHIA COLI CYTOCHROME B562
SOLUTION NMR
| NMR Experiment | ||||||||
|---|---|---|---|---|---|---|---|---|
| Experiment | Type | Sample Contents | Solvent | Ionic Strength | pH | Pressure | Temperature (K) | Spectrometer |
| 1 | 2D NOESY | 3MM R98C CYTOCHROME B562; 500MM PHOSPHATE BUFFER; 90% H2O, 10% D2O | 500mM PHOSPHATE | 4.8 | AMBIENT | 298 | ||
| 2 | 3D_15N-SEPARATED_NOESY | 2.5MM R98C CYTOCHROME B562; 500MM PHOSPHATE BUFFER; 90% H2O, 10% D2O | 500mM PHOSPHATE | 4.8 | AMBIENT | 298 | ||
| 3 | HNHA | 2.5MM R98C CYTOCHROME B562; 500MM PHOSPHATE BUFFER; 90% H2O, 10% D2O | 500mM PHOSPHATE | 4.8 | AMBIENT | 298 | ||
| 4 | 1D NOE | 3MM R98C CYTOCHROME B562; 500MM PHOSPHATE BUFFER; 90% H2O, 10% D2O | 500mM PHOSPHATE | 4.8 | AMBIENT | 298 | ||
| NMR Spectrometer Information | |||
|---|---|---|---|
| Spectrometer | Manufacturer | Model | Field Strength |
| 1 | Bruker | AVANCE | 800 |
| 2 | Bruker | AVANCE | 600 |
| NMR Refinement | ||
|---|---|---|
| Method | Details | Software |
| TORSION ANGLE DYNAMICS FOLLOWED BY ENERGY MINIMIZATION | A TOTAL OF 4325 NOESY CROSS-PEAKS WAS ASSIGNED, INTEGRATED AND TRANSFORMED IN UPPER DISTANCE LIMITS; 23 DISTANCE CONSTRAINTS WERE DERIVED FROM 1D NOE EXPERIMENTS INVOLVING FAST RELAXING PARAMAGNETIC SHIFTED SIGNALS. TOTALLY, THEY CORRESPONDED TO 2595 UPPER DISTANCE LIMITS, OF WHICH 2145 WERE FOUND TO BE MEANINGFUL. IN ADDITION, 45 3JHNHA COUPLINGS OBTAINED FROM THE HNHA 3D SPECTRUM AND 397 PCS WERE USED FOR THE STRUCTURE CALCULATIONS. | XwinNMR |
| NMR Ensemble Information | |
|---|---|
| Conformer Selection Criteria | ENERGY MINIMIZED AVERAGE STRUCTURE |
| Conformers Calculated Total Number | |
| Conformers Submitted Total Number | 1 |
| Representative Model | (minimized average structure) |
| Computation: NMR Software | ||||
|---|---|---|---|---|
| # | Classification | Version | Software Name | Author |
| 1 | processing | XwinNMR | ||
| 2 | structure solution | DYANA | 1.5 | GUENTERT P., MUMENTHALER, C., & WUTHRICH, K. |
| 3 | data analysis | XEASY | ECCLES, C., GUENTERT P., BILLETER, M., & WUTHRICH, K. | |
| 4 | structure solution | PSEUDODYANA | BANCI, L., BERTINI, I., CREMONINI, M.A., GORI SAVELLINI, G., LUCHINAT, C., WUTHRICH, K. & GUENTERT, P. | |
| 5 | refinement | Amber | 5.0 | PEARLMAN, D.A., CASE, D.A., CALDWELL, J.W., ROSS, W.S., CHEATHAM, T.E., FERGUSON, D.M., SEIBEL, G.L., SINGH, U.C., WEINER, P.K., & KOLLMAN, P.A. |
