3IOM
Crystal structure of Purine Nucleoside Phosphorylase from Mycobacterium tuberculosis in complex with 2'-Deoxyguanosine
Experimental procedure
Experimental method | SINGLE WAVELENGTH |
Source type | SYNCHROTRON |
Source details | LNLS BEAMLINE D03B-MX1 |
Synchrotron site | LNLS |
Beamline | D03B-MX1 |
Temperature [K] | 100 |
Detector technology | CCD |
Collection date | 2009-02-02 |
Detector | MAR CCD 165 mm |
Wavelength(s) | 1.42800 |
Spacegroup name | H 3 |
Unit cell lengths | 113.480, 113.480, 85.011 |
Unit cell angles | 90.00, 90.00, 120.00 |
Refinement procedure
Resolution | 28.340 - 2.140 |
R-factor | 0.1792 |
Rwork | 0.175 |
R-free | 0.24894 |
Structure solution method | MOLECULAR REPLACEMENT |
Starting model (for MR) | 1g2o |
RMSD bond length | 0.022 |
RMSD bond angle | 1.976 |
Data reduction software | MOSFLM |
Data scaling software | SCALA |
Phasing software | AMoRE |
Refinement software | REFMAC (5.2.0019) |
Data quality characteristics
Overall | Outer shell | |
Low resolution limit [Å] | 28.690 | 2.270 |
High resolution limit [Å] | 2.140 | 2.150 |
Rmerge | 0.086 | 0.138 |
Number of reflections | 22163 | |
<I/σ(I)> | 5.9 | 3.5 |
Completeness [%] | 96.9 | 81.8 |
Redundancy | 6.8 | 6.4 |
Crystallization Conditions
crystal ID | method | pH | temperature | details |
1 | VAPOR DIFFUSION, HANGING DROP | 7.6 | 293 | The protein was cocrystallized with a 1:0.5:1 stoichiometry of 500 mM Na2SO4 (Merck) and 3 mM 2dGuo (Fluka Biochemika), respectively, by hanging-drop vapor diffusion at 18 C. MtPNP protein (1 L of 25 mg mL-1 in 50 mM tris(hydroxymethyl)aminomethane (Tris) pH 7.6) had been previously mixed with an equal volume of the reservoir solution containing 100 mM Tris pH 8.0, 25% poly(ethylene glycol) (PEG) 3350, and 25 mM MgCl2 and hanging drops (3.5 L) were equilibrated against 400 L of the reservoir solution. , VAPOR DIFFUSION, HANGING DROP, temperature 293K |