3I7Z
Protein Tyrosine Phosphatase 1B - Transition state analog for the first catalytic step
Experimental procedure
Experimental method | SINGLE WAVELENGTH |
Source type | ROTATING ANODE |
Source details | RIGAKU RU200 |
Temperature [K] | 100 |
Detector technology | IMAGE PLATE |
Collection date | 2008-10-13 |
Detector | RIGAKU RAXIS IV |
Wavelength(s) | 1.5418 |
Spacegroup name | P 31 2 1 |
Unit cell lengths | 88.020, 88.020, 118.620 |
Unit cell angles | 90.00, 90.00, 120.00 |
Refinement procedure
Resolution | 27.637 - 2.300 |
R-factor | 0.207 |
Rwork | 0.206 |
R-free | 0.23300 |
Structure solution method | MOLECULAR REPLACEMENT |
Starting model (for MR) | 2cm2 |
RMSD bond length | 0.009 |
RMSD bond angle | 1.268 |
Data reduction software | d*TREK |
Data scaling software | d*TREK (9.4SSI) |
Phasing software | PHASER (1.3.3) |
Refinement software | PHENIX |
Data quality characteristics
Overall | Outer shell | |
Low resolution limit [Å] | 29.430 | 2.380 |
High resolution limit [Å] | 2.300 | 2.300 |
Rmerge | 0.075 | 0.540 |
Total number of observations | 13534 | |
Number of reflections | 24185 | |
<I/σ(I)> | 9.2 | 2.3 |
Completeness [%] | 99.9 | 100 |
Redundancy | 5.58 | 5.58 |
Crystallization Conditions
crystal ID | method | pH | temperature | details |
1 | VAPOR DIFFUSION, SITTING DROP | 7.5 | 277 | Drop: 2 uL of protein solution, 0.5 uL sucrose 30% (w/v) and 3 uL of precipitant solution (0.1 M HEPES pH 7.5, 0.2 M magnesium acetate and 15-17% polyethylene glycol 8000). Well: 500 uL of precipitant solution. The protein solution was prepared as follows: 0.36 uL of 100 mM of Na3VO4 and 10 uL of 50 mM of DADEYL peptide (at pH 8.5-9.0) were mixed and allowed to react for 1-1.5 hour; then, 50 uL of native PTP1B (12 mg/mL in 10 mM Tris pH 7.5, 25 mM NaCl, 0.2 mM EDTA and 3 mM DTT) was added and the solution used immediately for crystallization. VAPOR DIFFUSION, SITTING DROP, temperature 277K |