8TBR

Crystal Structure of Dihydrofolate reductase (DHFR) from Mycobacterium ulcerans Agy99 in complex with NADP and inhibitor MAM758

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SEALED TUBE
Source details	BRUKER D8 QUEST
Temperature [K]	100
Detector technology	PIXEL
Collection date	2023-06-26
Detector	Bruker PHOTON III
Wavelength(s)	1.5418
Spacegroup name	P 1 21 1
Unit cell lengths	28.606, 66.332, 43.973
Unit cell angles	90.00, 91.16, 90.00

Refinement procedure

Resolution	24.200 - 1.750
R-factor	0.163
Rwork	0.161
R-free	0.20370
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.010
RMSD bond angle	1.067
Data reduction software	XDS
Data scaling software	Aimless
Phasing software	PHASER
Refinement software	PHENIX (1.21rc1_4933)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	43.950	1.780
High resolution limit [Å]	1.750	1.750
Rmerge	0.118	1.404
Rmeas	0.123	1.504
Rpim	0.035	0.510
Total number of observations	205983	7701
Number of reflections	16609	911
<I/σ(I)>	18.1	1.5
Completeness [%]	99.9
Redundancy	12.4	8.5
CC(1/2)	0.997	0.480

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	6.5	291	Morpheus E4: 12.5%(v/v) MPD, 12.5%(v/v) PEG 1000, 12.5%(w/v) PEG 3350, 100 mM Imidazole/MES, pH 6.5, 30 mM Diethylene glycol, 30 mM Triethyleneglycol, 30 mM Tetraethylene glycol and 30 mM Pentaethylene glycol. MyulA.01062.a.B13.PS38720 at 8.9 mg/mL. 2mM MAM758 and 2mM NADP added to the protein prior to crystallization. Plate 13387 well E4 drop 2, Cryo: direct