1XPN
NMR structure of P. aeruginosa protein PA1324: Northeast Structural Genomics Consortium target PaP1
SOLUTION NMR
| NMR Experiment | ||||||||
|---|---|---|---|---|---|---|---|---|
| Experiment | Type | Sample Contents | Solvent | Ionic Strength | pH | Pressure | Temperature (K) | Spectrometer |
| 1 | 3D_15N-separated_NOESY | 1 mM U-13C,15N PA1324, 20 mM BisTrisPropane, 90% H2O, 10% D2O | 90% H2O/10% D2O | 300 mM NaCl | 6.8 | ambient | 298 | |
| 2 | 3D_13C-separated_NOESY | 1 mM U-13C,15N PA1324, 20 mM BisTrisPropane, 90% H2O, 10% D2O | 90% H2O/10% D2O | 300 mM NaCl | 6.8 | ambient | 298 | |
| 3 | 4D-13C,13C-HMQC-NOESY-HMQC | 1 mM U-13C,15N PA1324, 20 mM BisTrisPropane | 100% D2O | 300 mM NaCl | 6.8 | ambient | 298 | |
| NMR Spectrometer Information | |||
|---|---|---|---|
| Spectrometer | Manufacturer | Model | Field Strength |
| 1 | Varian | INOVA | 600 |
| 2 | Varian | INOVA | 750 |
| 3 | Varian | INOVA | 800 |
| NMR Refinement | ||
|---|---|---|
| Method | Details | Software |
| Molecular Dynamics and Simulated Annealing | NOE distance restraints were determined automatically using AutoStructure (G.T. Montelione & Y.J. Huang) starting from peak-picked NOESY data and chemical shift assignments. Final refinement was conducted in explicit solvent with Leonard-Jones and electrostatic potentials. Residues 1-26 are unrestrained and constitute a flexible, n-terminal tail. The structure of residues 27-170 is restrained by 1861 noe distance restraints (541 intraresidue, 513 sequential, 190 medium range, and 617 long range (n>4)), 132 dihedral restraints (65 phi, 67 psi), and 70 h-bond restraints (for 35 h-bonds). Dihedral restraints were derived from Talos and HNHA. H-bond restraints were derived from analysis of amides that exchange slowly with D2O solvent together with initial structures derived from Noesy data. | NIH-Xplor |
| NMR Ensemble Information | |
|---|---|
| Conformer Selection Criteria | structures with the lowest energy,structures with the fewest restraint violations |
| Conformers Calculated Total Number | 20 |
| Conformers Submitted Total Number | 20 |
| Representative Model | 1 (lowest energy, fewest violations, closest to the average) |
| Computation: NMR Software | ||||
|---|---|---|---|---|
| # | Classification | Version | Software Name | Author |
| 1 | refinement | NIH-Xplor | 2.0.4 | Brunger, Clore, Kuszewski, Schwieters, Tjandra |
| 2 | refinement | CNS | 1.1 | BRUNGER, ADAMS, CLORE, DELANO, GROS, GROSSE-KUNSTLEVE, JIANG, KUSZEWSKI, NILGES, PANNU, READ, RICE, SIMONSON, WARREN |
| 3 | data analysis | Sparky | 3.106 | T.D. Goddard, D.G. Kneller |
| 4 | structure solution | AutoStructure | 2.1.0 | G.T. Montelione, J.Y. Huang |
| 5 | data analysis | TALOS | 2003 | G. Cornilescu, F. Delaglio, A. Bax |
