8DSJ

Peptidylglycine alpha hydroxylating monooxygenase anaerobic

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	SSRL BEAMLINE BL12-2
Synchrotron site	SSRL
Beamline	BL12-2
Temperature [K]	100
Detector technology	PIXEL
Collection date	2021-10-27
Detector	DECTRIS PILATUS 6M
Wavelength(s)	1.000
Spacegroup name	P 1
Unit cell lengths	38.140, 53.263, 86.424
Unit cell angles	84.85, 89.96, 78.50

Refinement procedure

Resolution	37.400 - 2.800
R-factor	0.2012
Rwork	0.197
R-free	0.27490
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1phm
RMSD bond length	0.006
RMSD bond angle	1.472
Data reduction software	XDS
Data scaling software	Aimless (0.7.7)
Phasing software	MOLREP
Refinement software	REFMAC (5.8.0267)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	37.400	10.000	1.840
High resolution limit [Å]	1.800	8.000	1.800
Rmerge	0.169	0.162
Rmeas		0.172
Rpim		0.057
Number of reflections	57073	485	2118
<I/σ(I)>	3.6
Completeness [%]	92.8	98.8	57.5
Redundancy	7.9	8.8	4.8
CC(1/2)	0.855	0.982	0.328

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	4.5	298	1.5 uL of WT PHM protein at 17 mg/mL in 20 mM sodium phosphate, pH 7.5 was added to 1.5 uL of mother liquor solution containing 16-18% PEG 20K, 20-250 mM sodium citrate, and 2 mM CuSO4. Plates were sealed using transparent tape. Crystals were formed within one week, and these initial crystals were used to seed succeeding trays using the same crystal conditions. Seeding was performed using a Hampton Research seed bead and Hampton Research seeding tool. Initial crystals (5-7 crystals) were vortexed with seed beads for 30 seconds in 30 uL mother liquor, and streaked into a new drop using the seeding tool.