4C2B

Crystal Structure of High-Affinity von Willebrand Factor A1 domain with Disulfide Mutation in Complex with High Affinity GPIb alpha

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 22-ID
Synchrotron site	APS
Beamline	22-ID
Temperature [K]	93
Detector technology	CCD
Collection date	2013-04-05
Detector	MARRESEARCH MX-300
Spacegroup name	P 1 21 1
Unit cell lengths	96.530, 103.840, 119.930
Unit cell angles	90.00, 90.05, 90.00

Refinement procedure

Resolution	48.265 - 2.800
R-factor	0.2566
Rwork	0.253
R-free	0.28590
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	4c2a
RMSD bond length	0.002
RMSD bond angle	0.465
Data reduction software	XDS
Data scaling software	SCALEPACK
Refinement software	PHENIX ((PHENIX.REFINE))

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	48.300	2.870
High resolution limit [Å]	2.800	2.800
Rmerge	0.590
Number of reflections	56526
<I/σ(I)>	2.56	0.3
Completeness [%]	96.6	80.9
Redundancy	2.5

Crystallization Conditions

crystal ID	method	pH	temperature	details
1		4.6		CRYSTALS OF A1/SS-GPIB/VWD2 COMPLEX APPEARED IN DROPS WITH 20% PEG 4000, 0.16 M AMMONIUM SULFATE, 0.08M SODIUM ACETATE, PH 4.6, AND 20% GLYCEROL. THESE CRYSTALS WERE CRUSHED AND USED FOR SEEDING CRYSTAL GROWTH IN 8 MG/ML COMPLEX, 15% PEG 4000, 0.16 M AMMONIUM SULFATE, 0.08 M SODIUM ACETATE, PH 4.6, AND 20% GLYCEROL. SINCE THESE CRYSTALS WERE FORMED IN BUFFER CONTAINING 20% GLYCEROL, NO ADDITIONAL CRYOPROTECTION WAS USED.