8EKO

Sperm whale myoglobin mutant L29H F33W F43H (F33W CuBMb)

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ALS BEAMLINE 5.0.2
Synchrotron site	ALS
Beamline	5.0.2
Temperature [K]	100
Detector technology	PIXEL
Collection date	2019-06-14
Detector	DECTRIS PILATUS3 6M
Wavelength(s)	1.0
Spacegroup name	P 21 21 21
Unit cell lengths	40.059, 48.070, 77.739
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	35.610 - 1.340
R-factor	0.1406
Rwork	0.139
R-free	0.16420
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	4fwy
RMSD bond length	0.006
RMSD bond angle	0.959
Data reduction software	HKL-2000
Data scaling software	HKL-2000
Phasing software	PHENIX
Refinement software	PHENIX (1.19.2_4158)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	35.610	1.390
High resolution limit [Å]	1.340	1.340
Rmerge	0.192	0.057
Rmeas	0.027	0.080
Number of reflections	34356	3381
<I/σ(I)>	19.35
Completeness [%]	99.9	99.71
Redundancy	2	2
CC(1/2)	0.999	0.987

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	6.5	277	F33W CuBMb (1.0 mM) in 100 mM tris pH 8, (pH adjusted H2SO4), was mixed 1:3 with well buffer (0.1 M sodium cacodylate pH 6.5, 0.2 M sodium acetate trihydrate with 30% w/v polyethylene glycol 8000) using the hanging drop method with 300 uL well buffer in the well of the crystallization tray. Crystals obtained by this method were soaked with 10 eq CuSO4 in a mixture of F33W CuBMb and well buffer consistent with the original drop conditions to avoid dissolving the crystals and subsequently with 15eq potassium cyanide, in a mixture of E-F33W-CuBMb and well buffer using pH 7.0 sodium cacodylate