2XCU

Membrane-embedded monofunctional glycosyltransferase WaaA of Aquifex aeolicus, complex with CMP

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	MAX II BEAMLINE I911-2
Synchrotron site	MAX II
Beamline	I911-2
Temperature [K]	100
Detector technology	CCD
Collection date	2009-01-23
Detector	MARRESEARCH SX-165
Spacegroup name	P 1 21 1
Unit cell lengths	131.770, 44.670, 143.640
Unit cell angles	90.00, 97.15, 90.00

Refinement procedure

Resolution	31.220 - 2.420
R-factor	0.20478
Rwork	0.202
R-free	0.25087
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	2xci
RMSD bond length	0.015
RMSD bond angle	1.587
Data reduction software	MOSFLM
Data scaling software	SCALA
Phasing software	PHASER
Refinement software	REFMAC (5.5.0072)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	31.850	2.550
High resolution limit [Å]	2.420	2.420
Rmerge	0.040	0.140
Number of reflections	62789
<I/σ(I)>	21.3	6.8
Completeness [%]	97.2	88.3
Redundancy	3.6	3

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	8.5		HANGING-DROP VAPOR-DIFFUSION TECHNIQUE MIXING EQUAL VOLUMES OF PROTEIN SOLUTION (10-15 MG/ML, 25 MM TRIS-HCL, PH 8.7, 0.1 M NACL, 10% GLYCEROL, 2 MM CYMAL-6, 5 MM 2-MERCAPTOETHANOL) AND RESERVOIR (100 MM TRIS-HCL, PH 8.5, 35-40% (V/V) PEG 400, 200 MM NA-CITRATE, 50 MM 2-MERCAPTOETHANOL); SOAKING CRYSTALS IN 100 MM TRIS-HCL (PH 8.5), 36% (V/V) PEG 400, 200 MM NA-CITRATE AND 10 MM CMP FOR FOUR DAYS