6XG4

X-ray structure of Escherichia coli dihydrofolate reductase L28R mutant in complex with trimethoprim

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 19-ID
Synchrotron site	APS
Beamline	19-ID
Temperature [K]	100
Detector technology	PIXEL
Collection date	2018-07-12
Detector	DECTRIS PILATUS 6M
Wavelength(s)	0.97919
Spacegroup name	P 32 2 1
Unit cell lengths	61.548, 61.548, 104.661
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	37.370 - 2.100
R-factor	0.2493
Rwork	0.249
R-free	0.27790
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	WT DHFR solved in the same space group
RMSD bond length	0.006
RMSD bond angle	1.417
Data reduction software	HKL-3000
Data scaling software	HKL-3000
Phasing software	MOLREP
Refinement software	REFMAC (5.8.0267)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	50.000	2.120
High resolution limit [Å]	2.100	2.100
Rmeas	0.076	8.940
Rpim	0.025	3.012
Number of reflections	13880	334
<I/σ(I)>	37.5	0.7
Completeness [%]	99.9	100
Redundancy	9.3	8.8
CC(1/2)	1.000	0.672

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	5.6	293	0.1 M sodium citrate tribasic dihydrate (pH 5.6), 0.15 M ammonium acetate and 17.5% or 20% PEG 4000; 10 mM NADPH, 2 mM TMP was incubated with the L28R variant of DHFR overnight at 293 K