4KY8

Crystal structure of TS-DHFR from Cryptosporidium hominis in complex with NADPH, methotrexate, FdUMP and 4-((2-amino-6-methyl-4-oxo-4,7-dihydro-3H-pyrrolo[2,3-d]pyrimidin-5-yl)thio)-2-chlorophenyl)-L-glutamic acid

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	NSLS BEAMLINE X25
Synchrotron site	NSLS
Beamline	X25
Temperature [K]	77
Detector technology	PIXEL
Collection date	2013-04-16
Detector	DECTRIS PILATUS 6M
Wavelength(s)	1.10
Spacegroup name	C 1 2 1
Unit cell lengths	214.162, 115.665, 218.767
Unit cell angles	90.00, 94.36, 90.00

Refinement procedure

Resolution	48.120 - 3.084
R-factor	0.233
Rwork	0.232
R-free	0.26630
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1qzf
RMSD bond length	0.002
RMSD bond angle	0.543
Data reduction software	DENZO
Data scaling software	SCALEPACK
Phasing software	PHASER
Refinement software	PHENIX (1.8.1_1168)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	48.120	50.000	3.150
High resolution limit [Å]	3.080	8.400	3.100
Rmerge	0.207
Number of reflections	70173
<I/σ(I)>	4.1
Completeness [%]	71.6	99.8	11.3
Redundancy	3	3.3	1.3

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	8	298	14% (w/v) PEG 6000, 60 mM ammonium sulfate, 200 mM lithium sulfate and 100 mM Tris pH 8.0. ChTS-DHFR enzyme (~ 7 mg/ml) with 1 mM NADPH, 1 mM MTX, 1 mM FdUMP and 0.5 mM inhibitor., VAPOR DIFFUSION, HANGING DROP, temperature 298K