3INT

Structure of UDP-galactopyranose mutase bound to UDP-galactose (reduced)

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 21-ID-G
Synchrotron site	APS
Beamline	21-ID-G
Temperature [K]	100
Detector technology	CCD
Collection date	2008-11-05
Detector	MARMOSAIC 300 mm CCD
Wavelength(s)	0.97856
Spacegroup name	P 41
Unit cell lengths	94.078, 94.078, 129.443
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	30.000 - 2.510
R-factor	0.19643
Rwork	0.194
R-free	0.24720
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	3gf4
RMSD bond length	0.019
RMSD bond angle	1.771
Data reduction software	HKL-2000
Data scaling software	HKL-2000
Phasing software	MOLREP
Refinement software	REFMAC (5.5.0072)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	30.000	2.590
High resolution limit [Å]	2.500	2.500
Number of reflections	38510
<I/σ(I)>	15	6.7
Completeness [%]	100.0	100
Redundancy	7.6	7.5

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	5.6	298	Drops containing 1.5 microliters 5 mg/mL protein in 20 mM HEPES were combined with 1.5 microliters well solution (85 mM ammonium acetate, 42 mM tri-sodium citrate, 12.3% PEG 4000, 7.5% glycerol, 15 mM L-cysteine, 5 mM UDP-Glc) for 1-2 weeks. Crystals were then soaked in a solution of 53% Qiagen Cryos Suite Condition #87 with 15 mM L-cys, 30% methanol, 90 mM UDP-Galp (24hrs). Crystals were then soaked in a solution of 53% Qiagen Cryos Suite Condition #87 with 15 mM L-cys, 30% methanol, 90 mM UDP-Galp plus 100 mM sodium dithionite (3 min), pH 5.6, VAPOR DIFFUSION, HANGING DROP, temperature 298K