3BAM

RESTRICTION ENDONUCLEASE BAMHI COMPLEX WITH DNA AND MANGANESE IONS (POST-REACTIVE COMPLEX)

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	CHESS BEAMLINE F1
Synchrotron site	CHESS
Beamline	F1
Temperature [K]	110
Detector technology	IMAGE PLATE
Spacegroup name	P 21 21 21
Unit cell lengths	106.400, 79.600, 67.600
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	8.000 - 1.800
R-factor	0.226
Rwork	0.226
R-free	0.26800
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1bhm
RMSD bond length	0.010
RMSD bond angle	1.700
Data reduction software	DENZO
Data scaling software	SCALEPACK
Phasing software	X-PLOR
Refinement software	X-PLOR (3.851)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	40.000
High resolution limit [Å]	1.800
Rmerge	0.084	0.318 *
Total number of observations	111430 *
Number of reflections	40212
<I/σ(I)>	8.09
Completeness [%]	72.0	45.6 *
Redundancy	2.77

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	Vapor diffusion, hanging drop *	5.3	20 *	pH 5.3 CATALYSIS OCCURRED IN THE CRYSTAL AFTER SOAKING MANGANESE ION. THE ACTIVE SITE OF THE R SUBUNIT CLEAVED THE ORIGINAL D STRAND IN TWO (D AND E) RESULTING IN A POST-REACTIVE COMPLEX.

Crystallization Reagents in Literatures

ID	crystal ID	solution	reagent name	concentration (unit)	details
1	1	drop	protein	20-25 (mg/ml)
10	1	reservoir		0.15 (M)
11	1	reservoir	MES	0.1 (M)	pH5.3
12	1	reservoir	glycerol	5 (%)
2	1	drop		0.5 (M)
3	1	drop	potassium phosphate	20 (mM)	pH6.9
4	1	drop	dithiothreitol	1 (mM)
5	1	drop	glycerol	10 (%)
6	1	drop	DNA	10 (mg/ml)
7	1	drop	PEG8000	12-14 (%)
8	1	drop	MES	0.1 (M)	pH5.3
9	1	reservoir	PEG8000	12-14 (%)