5TX8
Solution structure of the de novo mini protein gHH_44
SOLUTION NMR
NMR Experiment | ||||||||
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Experiment | Type | Sample Contents | Solvent | Ionic Strength | pH | Pressure | Temperature (K) | Spectrometer |
1 | 3D C(CO)NH | 1 mM [U-99% 13C; U-99% 15N] HH2, 25 mM sodium acetate, 50 mM sodium chloride | 93% H2O/7% D2O | 0.075 M | 4.8 | 1 atm | 293 | Varian VXRS 600 |
2 | 3D 1H-15N NOESY | 1 mM [U-99% 13C; U-99% 15N] HH2, 25 mM sodium acetate, 50 mM sodium chloride | 93% H2O/7% D2O | 0.075 M | 4.8 | 1 atm | 293 | Varian INOVA 500 |
3 | 3D 1H-13C NOESY aliphatic | 1 mM [U-99% 13C; U-99% 15N] HH2, 25 mM sodium acetate, 50 mM sodium chloride | 93% H2O/7% D2O | 0.075 M | 4.8 | 1 atm | 293 | Varian VXRS 800 |
4 | 3D HNCA | 1 mM [U-99% 13C; U-99% 15N] HH2, 25 mM sodium acetate, 50 mM sodium chloride | 93% H2O/7% D2O | 0.075 M | 4.8 | 1 atm | 293 | Varian VXRS 800 |
5 | 2D 1H-13C HSQC aliphatic | 1 mM [U-99% 13C; U-99% 15N] HH2, 25 mM sodium acetate, 50 mM sodium chloride | 93% H2O/7% D2O | 0.075 M | 4.8 | 1 atm | 293 | Varian VXRS 800 |
6 | 2D 1H-15N HSQC | 1 mM [U-99% 13C; U-99% 15N] HH2, 25 mM sodium acetate, 50 mM sodium chloride | 93% H2O/7% D2O | 0.075 M | 4.8 | 1 atm | 293 | Varian VXRS 600 |
7 | D20 exchange | 1 mM [U-99% 13C; U-99% 15N] HH2, 25 mM sodium acetate, 50 mM sodium chloride | 100% D2O | 0.075 M | 4.8 | 1 atm | 293 | Varian VXRS 800 |
NMR Spectrometer Information | |||
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Spectrometer | Manufacturer | Model | Field Strength |
1 | Varian | VXRS | 600 |
2 | Varian | VXRS | 800 |
3 | Varian | INOVA | 500 |
NMR Refinement | ||
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Method | Details | Software |
torsion angle dynamics | STRUCTURE DETERMINATION WAS PERFORMED ITERATIVELY USING CYANA (AUTOMATED NOESY ASSIGNMENTS). A TOTAL OF 20 STRUCTURES OUT OF 100 WITH LOWEST TARGET FUNCTION FROM THE FINAL CYANA CALCULATION WERE TAKEN AND REFINED BY RESTRAINED MOLECULAR DYNAMICS/ENERGY MINIMIZATION IN EXPLICIT WATER AFTER ADDING 0% TO THE UPPER BOUNDARY LIMIT OF THE DISTANCE RESTRAINTS AND THE VDW LIMIT TO THE LOWER RESTRAINT. PARAM19 WAS USED FOR THE WATER REFINEMENT CALCULATIONS | CYANA |
NMR Ensemble Information | |
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Conformer Selection Criteria | target function |
Conformers Calculated Total Number | 100 |
Conformers Submitted Total Number | 20 |
Representative Model | 1 (closest to the average) |
Computation: NMR Software | ||||
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# | Classification | Version | Software Name | Author |
1 | refinement | CNS | 1.1 | Brunger A. T. et.al. |
2 | data analysis | Sparky | 3.115 | Goddard |
6 | processing | Felix | 2007 | Accelrys Software Inc. |
5 | structure calculation | CYANA | 2.1 | Guntert, Mumenthaler and Wuthrich |
3 | chemical shift assignment | PSVS | 1.5 | Bhattacharya and Montelione |