6ER4

Ruminococcus gnavus IT-sialidase CBM40 bound to alpha2,6 sialyllactose

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	ROTATING ANODE
Source details	RIGAKU MICROMAX-007 HF
Temperature [K]	100
Detector technology	CCD
Collection date	2014-07-17
Detector	RIGAKU SATURN 944+
Wavelength(s)	1.54178
Spacegroup name	P 1 21 1
Unit cell lengths	48.740, 72.200, 51.410
Unit cell angles	90.00, 103.88, 90.00

Refinement procedure

Resolution	49.910 - 1.300
R-factor	0.1097
Rwork	0.108
R-free	0.13450
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	2vw2
RMSD bond length	0.013
RMSD bond angle	1.712
Data reduction software	HKL-2000
Data scaling software	SCALA (3.3.21)
Phasing software	PHASER (2.5.6)
Refinement software	REFMAC (5.8.0103)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	49.910	49.909	1.340
High resolution limit [Å]	1.300	5.830	1.303
Rmerge		0.024	0.148
Rmeas	0.034
Rpim	0.013	0.008	0.143
Number of reflections	70604
<I/σ(I)>	32	23.2	5.5
Completeness [%]	83.9	99.5	13.6
Redundancy	5.1	10.1	1.4

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP		293	0.2M ammonium chloride with 20% PEG 8000 crystal growth time was improved using micro-seeding Crystals were cryoprotected using the crystallisation condition supplemented with 25% (w/w) glycerol crystal was co-crystallised with 10mM 2,6 sialyllactose and soaked with 100mM 2,6 sialyllactose immediately prior to freezing