2JIP

A New Catalytic Mechanism of Periplasmic Nitrate Reductase from Desulfovibrio desulfuricans ATCC 27774 from Crystallographic and EPR Data and based on detailed analysis of the sixth ligand

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ESRF BEAMLINE ID14-1
Synchrotron site	ESRF
Beamline	ID14-1
Temperature [K]	100
Detector technology	CCD
Collection date	2004-07-10
Detector	ADSC CCD
Spacegroup name	P 31 2 1
Unit cell lengths	106.317, 106.317, 134.032
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	92.060 - 2.300
R-factor	0.178
Rwork	0.175
R-free	0.25200
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	2nap
RMSD bond length	0.016
RMSD bond angle	1.739
Phasing software	REFMAC
Refinement software	REFMAC (5.2.0019)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	54.150	2.310
High resolution limit [Å]	2.190	2.190
Rmerge	0.110	0.990
Number of reflections	43422
<I/σ(I)>	11.2	1.1
Completeness [%]	95.7	71.1
Redundancy	5.9	4.1

Crystallization Conditions

crystal ID	method	pH	temperature	details
1		6		COCRYSTALLISATION WITH 200MM OF NO3 (NITRATE) IN 200MM OF DDNAPA (IE PROTEIN) WHICH HAD BEEN REDUCED BY DITHIONITE IN ANAEROBIC CONDITIONS AND THEN AIR REOXIDISED BEFORE SETTTING UP FOR CRYSTALLISATIONS STRAIGHT AFTER THAWING FROM 193K. SAMPLE WAS DIALYSED AND CONCENTRATED BEFORE CRYSTALLISING IN 0.1M MES PH 6.0, 7.5% PEG 8000