6F5M

Crystal structure of highly glycosylated human leukocyte elastase in complex with a thiazolidinedione inhibitor

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	2012-10-20
Detector	DECTRIS PILATUS 2M-F
Wavelength(s)	1.0000
Spacegroup name	H 3 2
Unit cell lengths	204.557, 204.557, 62.155
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	45.553 - 2.700
R-factor	0.1804
Rwork	0.176
R-free	0.23360
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1ppg
RMSD bond length	0.002
RMSD bond angle	0.542
Data reduction software	XDS
Data scaling software	Aimless
Phasing software	PHASER
Refinement software	PHENIX ((1.12_2829: ???))

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	45.600	2.830
High resolution limit [Å]	2.700	2.700
Rmerge	0.241
Number of reflections	13722	1808
<I/σ(I)>	15.4	1.2
Completeness [%]	99.9	100
Redundancy	20.7	21.6
CC(1/2)	0.998	0.442

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP		293	HLE from human blood was purchased from SERVA as a lyophilisate in the presence of sodium acetate puffer, pH 5.5. The lyophilisate was disolved in water so that the HLE concentration was 5 mg/ml (170 micromolar) and the acetate concentration 250 millimolar. 120 microliter dissolved HNE lysophilisate was mixed with 6 microliter inhibitor solution (10 mM in DMSO). 1 microliter of the resulting HLE/inhibitor mixture was mixed with 0.5 microliter reservoir solution which was composed of 20 % PEG MME 5000, 0.2 M potassium sulphate. Repeated seeding was necessary to get usable crystals. In the final seeding step the reservoir was composed of 20 % PEG MME 5000, 0.2 M sodium sulphate.