2V45

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 ID14-4
Synchrotron site	ESRF
Beamline	ID14-4
Temperature [K]	100
Detector technology	CCD
Collection date	2005-05-03
Detector	ADSC CCD
Spacegroup name	P 31 2 1
Unit cell lengths	105.902, 105.902, 130.241
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	91.670 - 2.400
R-factor	0.168
Rwork	0.165
R-free	0.24000
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	2nap
RMSD bond length	0.015
RMSD bond angle	1.727
Phasing software	PHASER
Refinement software	REFMAC (5.2.0019)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	65.090	2.530
High resolution limit [Å]	2.400	2.400
Rmerge	0.100	0.980
Number of reflections	33558
<I/σ(I)>	16.9	1.5
Completeness [%]	99.9	99.3
Redundancy	7	6.3

Crystallization Conditions

crystal ID	method	pH	temperature	details
1		6.5		COCRYSTALLISATION WITH 20MM OF KCLO4 (PERCHLORATE) IN 200MM OF DDNAPA (IE PROTEIN) WHICH HAD BEEN REDUCED BY DITHIONITE IN ANAEROBIC CONDITIONS AND THEN AIR REOXIDISED. THE SAMPLE WAS LEFT AT ROOM TEMPERATURE FOR SEVERAL MONTHS BEFORE SETTING UP CRYSTALLISATIONS IN 0.1M MES, PH 6.5, 8% PEG 8000