7SKM

Complex between S. aureus aureolysin and wt IMPI.

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ALBA BEAMLINE XALOC
Synchrotron site	ALBA
Beamline	XALOC
Temperature [K]	100
Detector technology	PIXEL
Collection date	2020-07-11
Detector	DECTRIS PILATUS 6M
Wavelength(s)	0.9793
Spacegroup name	P 41
Unit cell lengths	68.140, 68.140, 166.180
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	52.689 - 1.850
Rwork	0.164
R-free	0.21860
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1bqb 3ssb
RMSD bond length	0.008
RMSD bond angle	1.644
Data reduction software	XDS
Data scaling software	XSCALE
Phasing software	PHASER
Refinement software	REFMAC (5.8.0267)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	52.700	1.960
High resolution limit [Å]	1.850	1.850
Rmerge	0.149	2.772
Rpim	0.042	0.788
Number of reflections	64323	10209
<I/σ(I)>	14.7	1.9
Completeness [%]	100.0
Redundancy	13.6
CC(1/2)	0.999	0.630

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	5.5	293	Best crystals of aureolysin in complex with either IMPI variant were obtained at 20 degrees with protein solutions consisting of 5 mg/mL of aureolysin and 2.9 mg/mL of IMPI (peptidase:inhibitor molar ratio of 1:2.5) in 50 mM Tris-HCl, 150 mM sodium chloride, 1.6 mM calcium chloride, 8.3 microM zinc chloride, pH 8.0, which was mixed with reservoir solution comprising 0.1 M Bis-Tris, 25% (w/v) PEG 3350, pH 5.5.