6D45

L89S Mutant of FeBMb Sperm Whale Myoglobin

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	NSLS BEAMLINE X29A
Synchrotron site	NSLS
Beamline	X29A
Temperature [K]	100
Detector technology	CCD
Collection date	2015-12-08
Detector	ADSC QUANTUM 315
Wavelength(s)	0.83, 1.8
Spacegroup name	P 21 21 21
Unit cell lengths	39.359, 48.075, 78.237
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	35.160 - 1.779
R-factor	0.1682
Rwork	0.164
R-free	0.20530
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	3m38
RMSD bond length	0.006
RMSD bond angle	0.910
Data reduction software	HKL-2000
Data scaling software	HKL-2000
Phasing software	PHENIX (1.10.1_2155)
Refinement software	PHENIX (1.10.1_2155)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	50.000	50.000	1.810
High resolution limit [Å]	1.780	4.830	1.780
Rmerge	0.053	0.047	0.241
Rmeas	0.059	0.053	0.271
Rpim	0.026	0.024	0.121
Total number of observations	69715
Number of reflections	14785	818	613
<I/σ(I)>	18.5
Completeness [%]	99.1	97.6	86
Redundancy	4.7	4.2	4.6
CC(1/2)		0.996	0.950

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	6.57	277.15	Crystals of L89S-FeBMb were set up and grown at 4 C on hanging drops containing 0.1M MES pH 6.57, 0.2M NaOAC.3H2O, 30% PEG 6K as well buffer. Drops contained an equal volume of 1 mM L89S-FeBMb in 20 mM TRIS.H2SO4 pH 8 and the well buffer. Prior to mounting, the L89S-FeBMb crystals were soaked in buffer containing 0.1M MES pH 6.0, 0.2M NaOAC.3H2O, 30% PEG 6K for 30 min and then frozen in a cryoprotectant solution of 50 mM MES pH 6.0, and 30% PEG 400