4Q0D

Crystal structure of TS-DHFR from Cryptosporidium hominis in complex with NADPH, FdUMP, methotrexate and 2-amino-4-oxo-4,7-dihydro-pyrrolo[2,3-d]pyrimidine-methyl-phenyl-L-glutamic acid.

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	NSLS BEAMLINE X29A
Synchrotron site	NSLS
Beamline	X29A
Temperature [K]	77
Detector technology	CCD
Collection date	2014-03-07
Detector	ADSC QUANTUM 315r
Wavelength(s)	1.075
Spacegroup name	C 1 2 1
Unit cell lengths	213.611, 115.031, 217.643
Unit cell angles	90.00, 93.92, 90.00

Refinement procedure

Resolution	48.941 - 3.449
R-factor	0.2181
Rwork	0.217
R-free	0.24200
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	4ky8
RMSD bond length	0.002
RMSD bond angle	0.676
Data reduction software	DENZO
Data scaling software	SCALEPACK
Phasing software	PHASER
Refinement software	PHENIX (1.8.4_1496)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	50.000	50.000	3.510
High resolution limit [Å]	3.450	9.350	3.450
Rmerge	0.135	0.065	0.465
Number of reflections	62866
<I/σ(I)>	6.9
Completeness [%]	91.0	99.2	85.6
Redundancy	3.3	3.6	3

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	8	298	12% (w/v) PEG 6000, 60 mM ammonium sulfate, 200 mM lithium sulfate, and 100 mM Tris pH 8.0. ChTS-DHFR enzyme (approximately 7 mg/ml) with 1 mM NADPH, 1 mM FdUMP, 1 mM methotrexate and 0.5 mM inhibitor, VAPOR DIFFUSION, HANGING DROP, temperature 298K