4AHS

Parallel screening of a low molecular weight compound library: do differences in methodology affect hit identification

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	AUSTRALIAN SYNCHROTRON BEAMLINE MX1
Synchrotron site	Australian Synchrotron
Beamline	MX1
Temperature [K]	100
Detector technology	CCD
Collection date	2008-03-13
Detector	ADSC QUANTUM 210r
Spacegroup name	P 31
Unit cell lengths	71.183, 71.183, 66.653
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	61.660 - 1.750
R-factor	0.20763
Rwork	0.206
R-free	0.24787
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1bi4
RMSD bond length	0.022
RMSD bond angle	2.242
Data reduction software	MOSFLM
Data scaling software	SCALA
Phasing software	PHASER
Refinement software	REFMAC (5.6.0117)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	61.700	1.840
High resolution limit [Å]	1.750	1.750
Rmerge	0.070	0.450
Number of reflections	36824
<I/σ(I)>	16.2	1.9
Completeness [%]	96.5	80.9
Redundancy	4.9	2.6

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	5.5	293	PROTEIN: 5.5MG/ML IN 40MM TRIS BUFFER AT PH 8.0, 250MM NACL, 30MM MGCL2, 5MM DTT. CRYSTALLANT: 100MM SODIUM ACETATE PH 5.0 TO 5.5, 1.2 TO 1.5M AMMONIUM SULFATE AT 20C IN SITTING DROP PLATES. FRAGMENTS WERE SOAKED INTO PREFORMED CRYSTALS 24-48 HOURS PRIOR TO DATA COLLECTION.