4MXR

Crystal structure of Trypanosoma cruzi formiminoglutamase with Mn2+2

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	ROTATING ANODE
Source details	RIGAKU RU200
Temperature [K]	100
Detector technology	IMAGE PLATE
Collection date	2013-02-17
Detector	RIGAKU RAXIS IV++
Wavelength(s)	1.5418
Spacegroup name	H 3
Unit cell lengths	128.922, 128.922, 85.377
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	21.964 - 1.849
R-factor	0.1882
Rwork	0.186
R-free	0.21690
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	2a0m
RMSD bond length	0.004
RMSD bond angle	0.795
Data reduction software	HKL-2000
Data scaling software	HKL-2000
Phasing software	PHASER ((CCP4))
Refinement software	PHENIX ((phenix.refine: 1.5_2))

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	50.000	1.920
High resolution limit [Å]	1.850	1.850
Rmerge	0.059	0.438
Number of reflections	44906
<I/σ(I)>	14.162	2.013
Completeness [%]	99.2	98.8
Redundancy	2.5	2.4

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	4.9	294	A 4 UL drop of protein solution [10 mg/mL protein, 50 mM bicine (pH 8.5), 100 UM MnCl2, 1 mM TCEP] was mixed with a 4 UL drop of precipitant solution [31% PEG 300, 0.1 M sodium acetate (pH 4.9)] on a siliconized cover slide and equilibrated against a 500 UL reservoir of precipitant solution at 294K. The yielded crystal is apo. To obtain the Mn2+2 bound form, this apo-crystal was soaked with 5 mM MnCl2 in 0.1 M sodium malonate (pH 8.0), 27% PEG 3350) for 24 hours, VAPOR DIFFUSION, HANGING DROP