3CMX
Mechanism of homologous recombination from the RecA-ssDNA/dsDNA structures
Experimental procedure
Experimental method | SINGLE WAVELENGTH |
Source type | SYNCHROTRON |
Source details | APS BEAMLINE 24-ID-C |
Synchrotron site | APS |
Beamline | 24-ID-C |
Collection date | 2007-02-01 |
Spacegroup name | P 21 21 2 |
Unit cell lengths | 159.000, 300.500, 80.100 |
Unit cell angles | 90.00, 90.00, 90.00 |
Refinement procedure
Resolution | 40.000 - 3.400 |
R-factor | 0.239 |
Rwork | 0.238 |
R-free | 0.25600 |
Structure solution method | MOLECULAR REPLACEMENT |
RMSD bond length | 0.009 |
RMSD bond angle | 1.186 |
Refinement software | REFMAC (5.3.0036) |
Data quality characteristics
Overall | |
Low resolution limit [Å] | 40.000 |
High resolution limit [Å] | 3.400 |
Number of reflections | 49365 |
Crystallization Conditions
crystal ID | method | pH | temperature | details |
1 | 8 | RecA5 fusion protein was incubated with a 3-fold molar excess of ssDNA in the original protein buffer supplemented with 2 mM ADP, 10 mM MgCl2 and 8 mM AlF4, pH 6.0. Crystals of the RecA5-(ADP-AlF4-Mg)5-(dT)15 complex were grown from 50 mM Tris-Cl, 9% (w/v) PVP K15, 32% (v/v) MPD, 10 mM DTT, pH 8.0. RecA5-(ADP-AlF4-Mg)5-(dT)15-(dA)12 complex were obtained by soaking the RecA5-(ADP-AlF4-Mg)5-(dT)15 crystals in a 0.2 mM solution of the complementary (dA)12 oligonucleotide in 25 mM Tris-Cl, 9% (w/v) PVP K15, 32% (v/v) MPD, 2 mM ADP, 8 mM AlF4, and 10 mM MgCl2 for 4 hr. |
Crystallization Reagents
ID | crystal ID | solution ID | reagent name | concentration | details |
1 | 1 | 1 | ADP | ||
10 | 1 | 2 | MPD | ||
11 | 1 | 2 | DTT | ||
12 | 1 | 3 | 25 mM Tris-Cl | ||
13 | 1 | 3 | PVP K15 | ||
14 | 1 | 3 | MPD | ||
15 | 1 | 3 | ADP | ||
16 | 1 | 3 | AlF4 | ||
17 | 1 | 3 | MgCl2 | ||
2 | 1 | 1 | MgCl2 | ||
3 | 1 | 1 | AlF4 | ||
4 | 1 | 1 | 50 mM Tris-Cl | ||
5 | 1 | 1 | PVP K1 | ||
6 | 1 | 1 | MPD | ||
7 | 1 | 1 | DTT | ||
8 | 1 | 2 | 50 mM Tris-Cl | ||
9 | 1 | 2 | PVP K1 |