3UAN

Crystal structure of 3-O-sulfotransferase (3-OST-1) with bound PAP and heptasaccharide substrate

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	IMAGE PLATE
Collection date	2011-03-12
Detector	MAR scanner 345 mm plate
Wavelength(s)	1
Spacegroup name	P 1 21 1
Unit cell lengths	52.883, 62.168, 86.373
Unit cell angles	90.00, 95.16, 90.00

Refinement procedure

Resolution	24.570 - 1.844
R-factor	0.175
Rwork	0.174
R-free	0.20230
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1zrh
RMSD bond length	0.004
RMSD bond angle	0.838
Data reduction software	HKL-2000
Data scaling software	HKL-2000
Phasing software	MOLREP (Suite)
Refinement software	PHENIX ((phenix.refine: 1.7_650))

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	25.000	1.880
High resolution limit [Å]	1.844	1.844
Number of reflections	44993
<I/σ(I)>	21.8	3.11
Completeness [%]	97.0	84.6
Redundancy	3.7	2.9

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	MICROBATCH	5.5	293	Crystals of the 3-OST-1/PAP/heptasaccharide complex were obtained using micro-batch technique, by mixing 1.5 mL of the complex (12.9 mg/mL 3-OST-1, 5 mM heptasaccharide, 4 mM PAP, 23.6 mM Tris pH 7.5, 142 mM NaCl) was mixed with 2.5 mL of 0.1 M sodium citrate pH 5.5 and 20% PEG 3000. Crystals grew to a usable size after 10 days incubation at room temperature. The crystals were transferred in two steps to a cryoprotectant solution containing 0.1 M sodium citrate pH 5.5, 0.1 M NaCl, 4 mM PAP, 20 mM heptasaccharide, 30% PEG3000, and 7.6% ethylene glycol. , temperature 293K