1M0D

Crystal Structure of Bacteriophage T7 Endonuclease I with a Wild-Type Active Site and Bound Manganese Ions

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ESRF BEAMLINE ID14-2
Synchrotron site	ESRF
Beamline	ID14-2
Temperature [K]	100
Detector technology	CCD
Collection date	2001-06-25
Detector	ADSC QUANTUM 4
Wavelength(s)	0.933
Spacegroup name	P 21 21 2
Unit cell lengths	123.810, 133.960, 61.460
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	24.800 - 1.900
R-factor	0.21
Rwork	0.210
R-free	0.23400
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1fzr
RMSD bond length	0.008
RMSD bond angle	1.380 *
Data reduction software	MOSFLM
Data scaling software	CCP4 ((SCALA))
Phasing software	CNS
Refinement software	CNS

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	25.550	1.990
High resolution limit [Å]	1.900	1.900
Rmerge	0.085 *
Total number of observations	419157 *
Number of reflections	81257
<I/σ(I)>	7.6	1.6
Completeness [%]	99.9	99.4
Redundancy	5.2	4.4

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	Vapor diffusion, hanging drop *	8 *	291	PEG 4000, Ammonium sulphate, Sodium chloride, Tris HCL, pH 7.2, Vapor diffusion, hanging drop. Seeds of E65K mutant used, temperature 291K

Crystallization Reagents in Literatures

ID	crystal ID	solution	reagent name	concentration (unit)	details
1	1	drop	Tris-HCl	20 (mM)	pH8.0
2	1	drop		50 (mM)
3	1	drop	protein	4 (mg/ml)
4	1	reservoir	PEG4000	17-19 (%)
5	1	reservoir	ammonium sulfate	100 (mM)
6	1	reservoir	Tris-HCl	100 (mM)	pH7.2