5FC9

Novel Purple Cupredoxin from Nitrosopumilus maritimus

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	NSLS BEAMLINE X29A
Synchrotron site	NSLS
Beamline	X29A
Temperature [K]	80
Detector technology	CCD
Collection date	2011-04-27
Detector	ADSC QUANTUM 315r
Wavelength(s)	1.075
Spacegroup name	P 41 21 2
Unit cell lengths	68.887, 68.887, 184.450
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	45.870 - 1.600
R-factor	0.174
Rwork	0.173
R-free	0.19780
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	4azu
RMSD bond length	0.006
RMSD bond angle	1.068
Data reduction software	HKL-2000
Data scaling software	HKL-2000
Phasing software	PHENIX
Refinement software	PHENIX (1.9_1692)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	50.000	50.000	1.660
High resolution limit [Å]	1.600	3.450	1.600
Rmerge	0.105	0.087	0.719
Total number of observations	1760680
Number of reflections	59707
<I/σ(I)>	11
Completeness [%]	100.0	99.9	100
Redundancy	29.5	39.2	28.1

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	8	277	Crystals of N. mar_1307 were grown by the hanging drop vapor diffusion method. Protein solution was prepared in 50 mM sodium acetate buffer at pH 6.0 to a concentration of about 2 mM. A 2 uL portion of this protein solution was then mixed with 2 uL of a well buffer solution consisting of 0.1 M TrisHCl at pH 8.0, 20mM CuSO4, 0.1M LiNO3 and varying amounts of polyethylene glycol (PEG). The highest quality crystals formed from wells with 35 % w/v PEG 4000 after about 2 months at 4 C