4GFD

Thymidylate kinase (TMK) from S. Aureus in complex with TK-666

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 31-ID
Synchrotron site	APS
Beamline	31-ID
Temperature [K]	140
Detector technology	CCD
Collection date	2011-07-25
Detector	RAYONIX MX225HE
Wavelength(s)	0.97
Spacegroup name	P 1 21 1
Unit cell lengths	46.100, 90.540, 48.200
Unit cell angles	90.00, 103.08, 90.00

Refinement procedure

Resolution	27.880 - 1.800
R-factor	0.1694
Rwork	0.168
R-free	0.19750
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.010
RMSD bond angle	1.070
Data reduction software	PROCESS
Data scaling software	SCALA
Phasing software	AMoRE
Refinement software	BUSTER (2.11.2)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	90.060	1.540
High resolution limit [Å]	1.460	1.460
Rmerge	0.041	0.480
Number of reflections	65022
<I/σ(I)>	14	2.6
Completeness [%]	98.1	98.4
Redundancy	3.7	3.7

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP		293	Protein buffer: 25 mM Tris/HCl, pH 8.0, 0.15 M NaCl, 1 mM DTT, 1 mM EDTA, 20% Glycerol Well solution: 100 mM PCPT (propionate-cacodylate-bistris propane buffer) pH 7-8, 21-24% PEG 3350, 200 mM MgCl2, using 1:1 protein:reservoir solution with the protein solution at 13 mg/mL. Crystals were harvested and soaked overnight in a solution containing 100 mM PCPT, 35% PEG 3350, 200 mM Mg2Cl and 1-2 mM TK-666 from a 100 mM DMSO stock. After soaking the crystals were cryoprotected by soaking for 15 minutes in compound-soak solution supplemented with 20% ethylene glycol. , VAPOR DIFFUSION, SITTING DROP, temperature 293K