2IZO
Structure of an Archaeal PCNA1-PCNA2-FEN1 Complex
Experimental procedure
Experimental method | SINGLE WAVELENGTH |
Source type | SYNCHROTRON |
Source details | ESRF BEAMLINE ID29 |
Synchrotron site | ESRF |
Beamline | ID29 |
Temperature [K] | 100 |
Detector technology | CCD |
Collection date | 2005-09-07 |
Detector | ADSC CCD |
Spacegroup name | P 21 21 21 |
Unit cell lengths | 93.986, 99.774, 99.958 |
Unit cell angles | 90.00, 90.00, 90.00 |
Refinement procedure
Resolution | 30.000 - 2.900 |
R-factor | 0.253 |
Rwork | 0.250 |
R-free | 0.31200 |
Structure solution method | MOLECULAR REPLACEMENT |
Starting model (for MR) | PDB ENTRIES 1B43 FOR FEN1 AND 1VYJ FOR PCNA1 AND PCNA2 |
RMSD bond length | 0.007 |
RMSD bond angle | 1.031 |
Data reduction software | MOSFLM |
Data scaling software | SCALA |
Phasing software | PHASER |
Refinement software | REFMAC (5.2.0019) |
Data quality characteristics
Overall | Outer shell | |
Low resolution limit [Å] | 60.000 | 2.950 |
High resolution limit [Å] | 2.800 | 2.800 |
Rmerge | 0.080 | 0.570 |
Number of reflections | 23534 | |
<I/σ(I)> | 15.3 | 3 |
Completeness [%] | 99.3 | 99.8 |
Redundancy | 4.8 | 4.9 |
Crystallization Conditions
crystal ID | method | pH | temperature | details |
1 | VAPOR DIFFUSION, HANGING DROP | PRIOR TO CRYSTALLISATION THE PCNA1-PCNA2-PCNA3 COMPLEX WAS MIXED WITH FEN1 AT AN EQUIMOLAR RATIO AND INCUBATED FOR 20 MIN AT 294 K. PCNA1-PCNA2-FEN1 CO-CRYSTALS WERE GROWN BY THE VAPOUR DIFFUSION METHOD IN HANGING DROPS. PCNA3 WAS NOT PRESENT IN THE CRYSTAL LATTICE. DROPS WERE PREPARED BY MIXING 100 MM PROTEIN COMPLEX IN 20 MM TRIS-HCL PH 8.0, 200 MM NACL, 5 MM MGCL2, 2 MM DTT BUFFER SOLUTION WITH EQUAL VOLUMES OF 0.1M ACETATE PH 4.8, 8% PEG 8,000, 220 MM ZNOAC2 AND 30 MM GLYCYL-GLYCYL-GLYCINE. |