8A8K

PAP phosphatase from Methanothermococcus thermolithotrophicus refined to 3.1 A

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	SLS BEAMLINE X06DA
Synchrotron site	SLS
Beamline	X06DA
Temperature [K]	100
Detector technology	PIXEL
Collection date	2020-03-27
Detector	DECTRIS PILATUS 2M-F
Wavelength(s)	1.64566
Spacegroup name	I 4
Unit cell lengths	174.054, 174.054, 183.803
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	57.790 - 3.100
R-factor	0.1874
Rwork	0.186
R-free	0.21960
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	alphafold model
Data reduction software	XDS
Data scaling software	SCALA
Phasing software	PHENIX
Refinement software	PHENIX (1.19.2_4158)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	126.380	3.270
High resolution limit [Å]	3.100	3.100
Rmerge	0.197	2.483
Rmeas	0.204	2.575
Rpim	0.056	0.680
Number of reflections	45681	7240
<I/σ(I)>	12.3	1.1
Completeness [%]	92.1	100
Redundancy	13.3	14.3
CC(1/2)	0.998	0.477

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	7.6	291	The PAP phosphatase from Methanothermococcus thermolithotrophicus was concentrated to 20 mg.ml-1 in 25 mM Tris/HCl pH 7.6, 10% v/v glycerol, 2 mM dithiothreitol, and 150 mM NaCl. The protein was co-crystallized with Tb-Xo4 (10 mM final concentration), MnCl2 (2 mM final) and 2 mM PAP. The Tb-Xo4 is a nucleating/phasing agent, which should increase the crystallization performance, however, in this case the same crystalline form has been obtained in absence of the compound and diffracted to similar resolution. Crystals were obtained by mixing 0.7 ul of the sample with 1.6 M tri-sodium citrate in a 96-Well MRC 2-drop crystallization plates in polystyrene (SWISSCI) containing 90 ul of crystallization solution in the reservoir. No cryoprotectant was used.