6SAZ

Cleaved human fetuin-b in complex with crayfish astacin

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	2018-11-11
Detector	DECTRIS PILATUS3 6M
Wavelength(s)	1.0032
Spacegroup name	P 61
Unit cell lengths	90.700, 90.700, 283.900
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	78.500 - 3.000
Rwork	0.185
R-free	0.24700
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	6ht9
Data reduction software	XDS
Data scaling software	XSCALE
Phasing software	PHASER
Refinement software	BUSTER-TNT

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	78.500	3.180
High resolution limit [Å]	3.000	3.000
Rmerge	0.184	1.565
Rmeas	0.195	1.660
Number of reflections	26287	4165
<I/σ(I)>	11.1
Completeness [%]	99.7	98.9
Redundancy	9.2	8.9
CC(1/2)	0.996	0.626

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP		293	Crystallization assays were set up following the sitting-drop vapor diffusion method at the joint IBMB/IRB Automated Crystallography Platform of Barcelona Science Park. A Tecan robot (Tecan Trading) was used to prepare reservoir solutions, and a Cartesian Microsys 4000 XL robot (Genomic Solutions) or a Phoenix nanodrop robot (Art Robbins Instruments) dispensed nanocrystallization drops on 96x2-well Swissci Polystyrene MRC Crystallization Plates (Molecular Dimensions). Plates were stored at 4 or 20 degrees in thermostatic crystal farms (Bruker AXS). The astacin-hFB complex only crystallized after incubating the inhibitor (at 7.5 mg/mL) with six-fold molar excess of the peptidase in 10 mM Tris-HCl, 140 mM sodium chloride, pH 6.8. Crystals were obtained at 20 degrees in 200 nL:100 nL drops with protein complex solution and 20 percent (w/v) polyethylene glycol 3,350, 0.2 M sodium tartrate dibasic as reservoir solution.