9O4T

RT XFEL structure of Soybean Lipoxygenase-1 in large unit-cell

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	FREE ELECTRON LASER
Source details	SLAC LCLS BEAMLINE MFX
Synchrotron site	SLAC LCLS
Beamline	MFX
Temperature [K]	291
Detector technology	CCD
Collection date	2022-12-12
Detector	RAYONIX MX340-HS
Wavelength(s)	1.31
Spacegroup name	P 1 21 1
Unit cell lengths	96.031, 94.536, 50.529
Unit cell angles	90.00, 91.18, 90.00

Refinement procedure

Resolution	19.590 - 1.950
R-factor	0.2131
Rwork	0.212
R-free	0.23480
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.003
RMSD bond angle	0.542
Data reduction software	cctbx.xfel
Data scaling software	cctbx.xfel.merge
Phasing software	PHENIX (1.21.1_5286)
Refinement software	PHENIX (1.21.1_5286)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	19.590	1.980
High resolution limit [Å]	1.950	1.950
Number of reflections	65618	4590
<I/σ(I)>	4.416
Completeness [%]	98.9	99.78
Redundancy	77.59	12.19
CC(1/2)	0.989	0.567

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	BATCH MODE	5.5	291	In a 2 mL Eppendorf Tube, 0.5 mL of precipitant (0.4M NaOAc pH=5.5, 16% PEG-3350) and 0.5 mL of SLO (10 mg/mL in 50 mM NaOAc, pH=5.0) was combined together with ~10uL of seed stock (see 2.2.1.) being added to the tube cap prior to mixing by inversion. The tube was then placed on a Thermo Fisher Tuber Revolver Rotator, set to 30 RPM. Samples were left to mix overnight. Subsequently, samples were mixed in a ~1:1 ratio with 18% hydroxyethylcellulose (Sigma-Aldrich PN-09368) dissolved in SLO precipitant (0.4M NaOAc pH=5.5, 16% PEG-3350)