8DSN

Peptidylglycine alpha hydroxylating monoxygenase, Q272A

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)	1.00
Spacegroup name	P 1
Unit cell lengths	38.878, 53.488, 86.056
Unit cell angles	84.71, 89.82, 77.96

Refinement procedure

Resolution	38.050 - 2.800
R-factor	0.2033
Rwork	0.198
R-free	0.29640
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1phm
RMSD bond length	0.008
RMSD bond angle	1.610
Data reduction software	XDS
Data scaling software	Aimless (0.7.7)
Phasing software	MOLREP
Refinement software	REFMAC (5.8.0267)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	38.050	2.050
High resolution limit [Å]	2.000	2.000
Rpim	0.438
Number of reflections	43540	3079
<I/σ(I)>	6
Completeness [%]	95.4	90.2
Redundancy	15.5	10.9
CC(1/2)		0.143

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.