8DSL

Peptidylglycine alpha hydroxylating monooxygenase, Q272E

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	2021-01-16
Detector	DECTRIS PILATUS3 6M
Wavelength(s)	0.979
Spacegroup name	P 1
Unit cell lengths	39.166, 53.535, 86.139
Unit cell angles	85.03, 89.79, 77.62

Refinement procedure

Resolution	38.250 - 2.050
R-factor	0.198
Rwork	0.196
R-free	0.23920
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1phm
RMSD bond length	0.010
RMSD bond angle	1.701
Data reduction software	XDS
Data scaling software	Aimless (0.7.4)
Phasing software	MOLREP
Refinement software	REFMAC (5.8.0267)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	38.250	2.110
High resolution limit [Å]	2.050	2.050
Rmerge	0.071	0.539
Rmeas	0.076	0.574
Rpim	0.025	0.194
Number of reflections	41882	3186
<I/σ(I)>	15.9	3.5
Completeness [%]	98.1	96.8
Redundancy	8.9	8.7
CC(1/2)	0.999	0.950

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	4.5	298	Hampton Research Cryschem 24-well plates. 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.