8QGX

Complex between human neutrophil elastase with nanobody NbE201

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	SOLEIL BEAMLINE PROXIMA 1
Synchrotron site	SOLEIL
Beamline	PROXIMA 1
Temperature [K]	293
Detector technology	PIXEL
Collection date	2022-02-12
Detector	DECTRIS EIGER X 16M
Wavelength(s)	0.9786
Spacegroup name	P 21 21 21
Unit cell lengths	64.980, 73.000, 84.920
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	48.540 - 2.300
R-factor	0.226
Rwork	0.224
R-free	0.27100
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.010
RMSD bond angle	1.130
Data reduction software	XDS
Data scaling software	XDS
Phasing software	PHASER
Refinement software	BUSTER (2.10.3)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	48.540	2.420
High resolution limit [Å]	2.300	2.300
Rmerge	0.128	1.914
Number of reflections	18530	2962
<I/σ(I)>	11.6
Completeness [%]	99.7
Redundancy	13.2
CC(1/2)	0.999	0.886

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP		293	The crystals were grown at 20oC using the sitting drop vapor diffusion method. The drop contained 0.2 uL of hNE in complex with NbE201 at a concentration of 15 mg/mL and 0.2 uL of 0.1M HEPES pH7 buffer with 10 per cent (v/v) polyethylene glycol 5000 monomethyl ether and 5 per cent (v/v) tacsimate. The crystals were transferred to a cryoprotectant solution containing 33 per cent (v/v) polyethylene glycol 6000 and 33 per cent (v/v) glycerol before being frozen in liquid nitrogen.