4O9K

Crystal structure of the CBS pair of a putative D-arabinose 5-phosphate isomerase from Methylococcus capsulatus in complex with CMP-Kdo

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 21-ID-F
Synchrotron site	APS
Beamline	21-ID-F
Temperature [K]	100
Detector technology	CCD
Collection date	2013-11-11
Detector	MARMOSAIC 225 mm CCD
Wavelength(s)	0.97872
Spacegroup name	P 21 21 21
Unit cell lengths	45.151, 62.408, 75.664
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	48.140 - 1.850
R-factor	0.1505
Rwork	0.148
R-free	0.18890
Structure solution method	SAD
RMSD bond length	0.016
RMSD bond angle	1.835
Data reduction software	HKL-3000
Data scaling software	SCALEPACK
Phasing software	SHELX
Refinement software	REFMAC (5.8.0049)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	50.000	1.880
High resolution limit [Å]	1.850	1.850
Rmerge	0.146	0.703
Number of reflections	18460
<I/σ(I)>	12.5	1.8
Completeness [%]	98.6	99.2
Redundancy	5.7	5.4

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	7.5	289	0.2 ul of 9 mg/ml protein in 20 mM HEPES pH 7.5, 150 mM NaCl, 10% Glycerol, 0.1% Sodium Azide and 0.5 mM TCEP were mixed with 0.2 ul of the MCSG-II condition #27 (0.2 M Ammomium Acetate, 0.1 M HEPES:NaOH pH 7.5, 25% (w/v) PEG 3350) and equilibrated against 1.5 M NaCl solution in QIAGEN EasyXtal 15-Well Tool plate. Before crystallization protein was incubated with 1/50 v/v of 2 mg/ml chymotrypsin solution at 289 K for 3 hours, VAPOR DIFFUSION, SITTING DROP