7BBM

Mutant nitrobindin M75L/H76L/Q96C/M148L (NB4H) from Arabidopsis thaliana with cofactor MnPPIX

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ESRF BEAMLINE ID30B
Synchrotron site	ESRF
Beamline	ID30B
Temperature [K]	100
Detector technology	PIXEL
Collection date	2020-09-17
Detector	DECTRIS PILATUS3 6M
Wavelength(s)	0.976253
Spacegroup name	P 21 21 2
Unit cell lengths	59.900, 79.547, 36.560
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	39.770 - 1.140
R-factor	0.1451
Rwork	0.144
R-free	0.15810
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	3wjb
RMSD bond length	0.012
RMSD bond angle	1.339
Data reduction software	XDS (20200417)
Data scaling software	Aimless (0.7.4)
Phasing software	PHASER (2.8.3)
Refinement software	PHENIX (1.19rc5_4047)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	39.774	39.774	1.157
High resolution limit [Å]	1.137	3.087	1.137
Rmerge	0.075	0.062	1.037
Rmeas	0.078	0.064	1.088
Rpim	0.022	0.018	0.325
Number of reflections	64928	3490	3190
<I/σ(I)>	14.5	37.595	2.164
Completeness [%]	99.9	99.77	100
Redundancy	12.2	11.72	10.93
CC(1/2)	0.998	0.997	0.811

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	MICROBATCH	8.6	285.15	1:1 mixture (2 uL total volume) of protein (20 mg/mL) and precipitant solution containing 100 mM Tris/HCl pH 8.6, 18 % (w/v) PEG 2000; 30% (v/v) Ethylene glycol were added as cryo-protectant before harvesting and flash freezing in liquid nitrogen.