4C2A

Crystal Structure of High-Affinity von Willebrand Factor A1 domain with R1306Q and I1309V Mutations in Complex with High Affinity GPIb alpha

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 23-BM-B
Synchrotron site	APS
Beamline	23-BM-B
Temperature [K]	93
Detector technology	CCD
Collection date	2012-10-26
Detector	MARRESEARCH
Spacegroup name	P 61
Unit cell lengths	89.060, 89.060, 124.520
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	48.446 - 2.081
R-factor	0.162
Rwork	0.161
R-free	0.18900
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1sq0
RMSD bond length	0.003
RMSD bond angle	0.688
Data reduction software	MOSFLM
Data scaling software	SCALEPACK
Refinement software	PHENIX ((PHENIX.REFINE: 1.8.2_1309))

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	48.400	2.190
High resolution limit [Å]	2.080	2.080
Rmerge	0.090	0.740
Number of reflections	66245
<I/σ(I)>	8.9	1.7
Completeness [%]	99.9	99.5
Redundancy	2.5

Crystallization Conditions

crystal ID	method	pH	temperature	details
1		6.5		CRYSTALS OF A1/VWD2-GPIBALPA/VWD2 COMPLEX APPEARED IN 12% PEG 8000, 0.1 M SODIUM CACODYLATE PH 6.5, 0.1 M CALCIUM ACETATE, AND 0.4 MG/ML RISTOCETIN (SIGMA-ALDRICH). THESE CRYSTALS WERE CRUSHED AND USED FOR SEEDING. CRYSTALS GREW IN 10 MG/ML COMPLEX, 14% PEG8000, 0.2 M CALCIUM ACETATE, 0.1 M SODIUM CACODYLATE, PH 6.5, AND 0.4 MG/ML RISTOCETIN. NEXT WE SOAKED THESE CRYSTALS IN 14% PEG8000, 0.2 M CALCIUM ACETATE, 0.1 SODIUM CACODYLATE, PH 6.5, AND 4 MG/ML RISTOCETIN.