7L3Y

Crystal structure of oxy-I107E CuB myoglobin (I107E L29H F43H sperm whale myoglobin; partial occupancy)

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	SSRL BEAMLINE BL11-1
Synchrotron site	SSRL
Beamline	BL11-1
Temperature [K]	100
Detector technology	PIXEL
Collection date	2015-07-20
Detector	DECTRIS PILATUS 6M
Wavelength(s)	0.98
Spacegroup name	P 21 21 21
Unit cell lengths	39.845, 46.982, 78.034
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	39.020 - 1.180
R-factor	0.1582
Rwork	0.157
R-free	0.19020
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	4fwx
RMSD bond length	0.013
RMSD bond angle	1.397
Data reduction software	XDS (v2015-03-01)
Data scaling software	Aimless (0.5.12)
Phasing software	PHENIX (1.10.1_2155)
Refinement software	PHENIX (1.19rc7_4070)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	39.020	39.020	1.200
High resolution limit [Å]	1.180	6.470	1.180
Rmerge	0.024	0.017	0.216
Rmeas	0.028	0.019	0.275
Rpim	0.013	0.009	0.169
Total number of observations	182510	1448	3107
Number of reflections	47352	360	1510
<I/σ(I)>	31.7	92.6	4.1
Completeness [%]	97.0	99.1	63.4
Redundancy	3.9	4	2.1
CC(1/2)	1.000	0.999	0.902

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	8.6	277	1.5-2.0 mM protein in 20 mM tris sulfate pH 8.0 buffer, mixed 1:1 with crystallization solution composed of 100 mM tris sulfate pH 8.6, 200 mM sodium acetate, and 30% w/v PEG 10000. After crystalization, soaked, under anaerobic conditions, in 20 mM dithionite for 10 minutes, and washed with anaerobic buffer. Reduced crystals were sealed in an air-tight bomb and pressurized at ~1500 psi O2 for up to 10 minutes. Crystals were harvested and cryocooled after a brief soak in 50% PEG 400 supersaturated with O2.