1T5R

STRUCTURE OF THE PANTON-VALENTINE LEUCOCIDIN S COMPONENT FROM STAPHYLOCOCCUS AUREUS

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	2002-11-08
Detector	ADSC QUANTUM 4
Wavelength(s)	0.9340
Spacegroup name	P 43
Unit cell lengths	94.850, 94.850, 306.110
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	35.500 - 2.000
Rwork	0.209
R-free	0.24700
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1pvl
RMSD bond length	0.006
RMSD bond angle	1.300
Data reduction software	MOSFLM
Data scaling software	CCP4 ((SCALA))
Phasing software	MOLREP
Refinement software	CNS (1.0)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	35.500	2.110
High resolution limit [Å]	2.000	2.000
Number of reflections	169450
<I/σ(I)>	9.1	1.6
Completeness [%]	93.6	67.5
Redundancy	5.9	2.5

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	8	285	CRYSTALLIZATION CONDITIONS: VAPOR EQUILIBRIUM USING THE HANGING METHOD AT 285K. THE PROTEINS WERE CONCENTRATED AT 20-25 MG/ML IN MES-NAOH PH 6.0 (50MM), NACL (50MM). DROPS WERE PREPARED BY MIXING EQUAL VOLUMES OF PROTEIN AND RESERVOIR SOLUTIONS AND USING RESERVOIR VOLUMES OF 500 microL CONTAINING 30% JEFFAMINE M-600, 0.1 M TRIS-HCL AT PH VALUES RANGING FROM 8.0 TO 8.9. MICRO- AND MACRO-SEEDING TECHNIQUES, INCLUDING CROSS-SEEDINGS, WERE USED TO GROW CRYSTALS. CRYSTAL SIZE (200X200X200 MM3) IN 6-10microL HANGING DROPS. , VAPOR DIFFUSION, HANGING DROP