8ESU

Myoglobin variant Mb-imi complex

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 24-ID-C
Synchrotron site	APS
Beamline	24-ID-C
Temperature [K]	100
Detector technology	PIXEL
Collection date	2019-11-01
Detector	DECTRIS PILATUS 6M-F
Wavelength(s)	0.9791
Spacegroup name	P 6
Unit cell lengths	90.425, 90.425, 45.442
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	78.310 - 1.040
R-factor	0.129
Rwork	0.128
R-free	0.14060
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	6m8f
RMSD bond length	0.008
RMSD bond angle	1.019
Data reduction software	XDS
Data scaling software	Aimless
Phasing software	PHENIX
Refinement software	PHENIX (1.20.1_4487)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	78.310	1.060
High resolution limit [Å]	1.040	1.040
Rmerge	0.067	0.504
Rpim	0.022	0.234
Number of reflections	101447	4642
<I/σ(I)>	18.2	2.7
Completeness [%]	99.6
Redundancy	9.2
CC(1/2)	0.995	0.817

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION		296	Mb(H64G,V68A) at 3.6 mM in 20 mM Tris-HCl pH 8.4, 1 mM EDTA, was mixed with an equal volume of reservoir solution (200 mM Tris-HCl pH 8.9 and 2.46 M ammonium sulfate). The crystal was dunked in a cryoprotection solution (reservoir buffer supplemented with 9% sucrose (w/v), 2% glucose (w/v), 8% glycerol (v/v), and 8% ethylene glycol (v/v)) prior to being flash-frozen in liquid nitrogen. Note that imidazole was present in lysis buffer at 10 mM prior to two-step purification.