8A0M

Capsular polysaccharide synthesis multienzyme in complex with capsular polymer fragment

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	DIAMOND BEAMLINE I04
Synchrotron site	Diamond
Beamline	I04
Temperature [K]	100
Detector technology	PIXEL
Collection date	2022-02-26
Detector	DECTRIS EIGER2 XE 16M
Wavelength(s)	0.9795
Spacegroup name	P 1 21 1
Unit cell lengths	127.911, 232.806, 129.177
Unit cell angles	90.00, 118.39, 90.00

Refinement procedure

Resolution	63.940 - 3.600
R-factor	0.2222
Rwork	0.220
R-free	0.26100
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	Alphafold
RMSD bond length	0.003
RMSD bond angle	0.567
Data reduction software	xia2 (dials)
Data scaling software	xia2 (dials)
Phasing software	PHASER
Refinement software	PHENIX (1.19.2_4158)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	63.940	3.729
High resolution limit [Å]	3.600	3.600
Rmeas	0.183	0.884
Number of reflections	76828	7658
<I/σ(I)>	8.79	1.38
Completeness [%]	99.7	99.75
Redundancy	7.1	6.7
CC(1/2)	0.988	0.796

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP		291	0.06M Magnesium chloride hexahydrate, 0.06M Calcium chloride dihydrate, 0.1M Imidazole, 0.1M MES monohydrate (acid) pH 6.5, MPD, PEG 1000, PEG 3350 (37.5%(v/v)) protein (9.5mg/ml) and purified polymer (produced by the multienzyme in-vitro)