9CRZ

Acanthamoeba Polyphaga Mimivirus R655

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 22-ID
Synchrotron site	APS
Beamline	22-ID
Temperature [K]	110
Detector technology	PIXEL
Collection date	2022-10-18
Detector	DECTRIS EIGER X 16M
Wavelength(s)	1.0
Spacegroup name	P 21 21 21
Unit cell lengths	60.634, 96.213, 133.454
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	78.167 - 2.100
Rwork	0.210
R-free	0.25180
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.004
RMSD bond angle	1.023
Data reduction software	xia2 (3.8.0)
Data scaling software	Aimless (0.7.8)
Phasing software	PHENIX (1.20.1)
Refinement software	REFMAC (5.8.0425)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	78.170	2.160
High resolution limit [Å]	2.100	2.100
Number of reflections	46085	3706
<I/σ(I)>	6.3	1.7
Completeness [%]	99.6	99.5
Redundancy	5.1	5.2
CC(1/2)	0.946	0.706

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP		277	R655 IMAC elution fractions were digested with PreScission, and the His6 tag was removed by subtraction IMAC. Subtraction IMAC elution fractions were combined and concentrated for crystal trials. Single crystals were obtained through hanging drop vapor diffusion with the Mosquito liquid handling robot (TTPLabtech), utilizing a 200-nL drop with 1:1 mixture of protein solution and mother liquor at 277K. The protein solution has R655 (30 mg/mL) supplemented with 2 mM manganese(II) chloride and 2 mM UDP glucose. The mother liquor has 0.2 M magnesium formate, 20% (w/v) PEG 3350. To harvest the crystals, the crystals were soaked in a 1:1 mixture of 50% glycerol and mother liquor for 2 minutes and cooled in liquid nitrogen.