3H4W

Structure of a Ca+2 dependent Phosphatidylinositol-specific phospholipase C (PI-PLC) Enzyme from Streptomyces antibioticus

Experimental procedure

Experimental method	SAD
Source type	SYNCHROTRON
Source details	SSRL BEAMLINE BL9-2
Synchrotron site	SSRL
Beamline	BL9-2
Temperature [K]	100
Detector technology	CCD
Collection date	2008-01-01
Detector	MARMOSAIC 325 mm CCD
Wavelength(s)	0.9796
Spacegroup name	P 21 21 2
Unit cell lengths	52.377, 155.172, 41.409
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	77.590 - 1.500
R-factor	0.185
Rwork	0.184
R-free	0.20000
Structure solution method	SAD
RMSD bond length	0.005
RMSD bond angle	0.995
Data reduction software	MOSFLM
Data scaling software	SCALA (CCP4_3.2.25)
Phasing software	SHELX
Refinement software	REFMAC (refmac_5.5.0068)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	77.610	1.580
High resolution limit [Å]	1.500	1.500
Rmerge	0.058	0.220
Total number of observations		36537
Number of reflections	52171
<I/σ(I)>	8.2689	3.19
Completeness [%]	95.2	81.66
Redundancy	11.65	5.76

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	4.6	277	21% PEG 10000, 85mM sodium acetate pH 4.6, 170mM ammonium acetate, 9% glycerol. SeMe-derived crystals obtained with initial microseeding with underivatized crystals followed by a series of macroseedings with derivatized enzyme and same conditions., VAPOR DIFFUSION, HANGING DROP, temperature 277K