1H29

Sulfate respiration in Desulfovibrio vulgaris Hildenborough: Structure of the 16-heme Cytochrome c HmcA at 2.5 A resolution and a view of its role in transmembrane electron transfer

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ESRF BEAMLINE ID29
Synchrotron site	ESRF
Beamline	ID29
Temperature [K]	100
Detector technology	CCD
Collection date	2001-06-15
Detector	ADSC QUANTUM 4r
Spacegroup name	P 62
Unit cell lengths	220.390, 220.390, 102.640
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	25.000 * - 2.510
R-factor	0.195
Rwork	0.192
R-free	0.25800
Structure solution method	MAD
RMSD bond length	0.014
RMSD bond angle	1.550 *
Data reduction software	DENZO
Data scaling software	SCALEPACK
Refinement software	REFMAC (5.1.24)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	25.000	2.590
High resolution limit [Å]	2.500	2.500
Rmerge	0.064	0.354
Number of reflections	89606
<I/σ(I)>	11.9	3
Completeness [%]	91.3	84
Redundancy	6.5

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	Microdialysis *	7.5	277	EACH CRYSTALLIZATION ASSAY WAS PREPARED BY ADDING 5 UL OF THE PROTEIN SOLUTION (10 MG/ML) TO A DIALYSIS BUTTON WHICH WAS THEN COMPLETELY SOAKED IN THE CRYSTALLIZATION WAS THEN COMPLETELY SOAKED IN THE SOLUTION. AFTER A FEW DAYS AT 4C, SMALL HEXAGONAL BIPYRAMIDS STARTED STARTED TO APPEAR AND REACHED THEIR MAXIMUM SIZE AFTER ONE WEEK. THE BEST CRYOCRYSTALLOGRAPHY CONDITIONS WERE OBTAINED BY SOAKING THESE CRYSTALS IN A CRYSTALLIZATION SOLUTION CONTAINING 7.5% MPD, pH 7.50

Crystallization Reagents in Literatures

ID	crystal ID	solution	reagent name	concentration (unit)	details
1	1	reservoir	2-propanol	40 (%(v/v))
2	1	reservoir	Tris-HCl	0.1 (M)	pH7.5
3	1	drop	protein	10 (mg/ml)