4MYN

Crystal structure of Trypanosoma cruzi formiminoglutamase N114H variant with Mn2+2

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	NSLS BEAMLINE X29A
Synchrotron site	NSLS
Beamline	X29A
Temperature [K]	100
Detector technology	CCD
Collection date	2012-10-19
Detector	ADSC QUANTUM 315r
Wavelength(s)	1.075
Spacegroup name	H 3
Unit cell lengths	129.431, 129.431, 42.522
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	30.012 - 1.799
R-factor	0.1897
Rwork	0.188
R-free	0.22150
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	2a0m
RMSD bond length	0.007
RMSD bond angle	1.025
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	1.860
High resolution limit [Å]	1.800	1.800
Rmerge	0.088	0.754
Number of reflections	24658
<I/σ(I)>	18.949	2.966
Completeness [%]	100.0	100
Redundancy	6.3	6.1

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	7.5	294	Apo-enzyme was crystallized in the following conditions: a 4 uL drop of protein solution [10 mg/mL protein, 50 mM bicine (pH 8.5), 100 uM MnCl2] was mixed with a 4 uL drop of precipitant solution [25% PEG 3350, 0.1 M sodium acetate (pH 4.6)] on a siliconized cover slide and equilibrated against a 500 uL reservoir of precipitant solution. To obtain the Mn2+ bound form, apo-crystal was soaked in 10 mM MnCl2, 0.1 M sodium malonate (pH 7.5), 27% PEG 3350 for 24 hours, VAPOR DIFFUSION, SITTING DROP, temperature 294K