9EVO

Plasmodium falciparum apical membrane antigen 3D7

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	SOLEIL BEAMLINE PROXIMA 1
Synchrotron site	SOLEIL
Beamline	PROXIMA 1
Temperature [K]	100
Detector technology	PIXEL
Collection date	2014-03-21
Detector	DECTRIS PILATUS 6M
Wavelength(s)	0.97857
Spacegroup name	P 1
Unit cell lengths	38.230, 62.060, 71.500
Unit cell angles	89.13, 89.90, 83.92

Refinement procedure

Resolution	46.370 - 2.100
R-factor	0.20977
Rwork	0.207
R-free	0.25599
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.008
RMSD bond angle	1.729
Data reduction software	XDS
Data scaling software	SCALA
Phasing software	PHASER
Refinement software	REFMAC (5.8.0415)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	47.060	2.210
High resolution limit [Å]	2.100	2.100
Rmerge	0.199	0.872
Rmeas	0.268	1.183
Rpim	0.179	0.795
Number of reflections	37352	5400
<I/σ(I)>	4.3	2.1
Completeness [%]	97.9
Redundancy	2
CC(1/2)	0.902	0.215

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP		290	Crystals of PfAMA1-3D7 were obtained by mixing 1.2 micro-L of protein and 1.2 micro-L of reservoir buffer containing 12% PEG 3350, 0.1M Hepes pH 7 and 10% propanol-2. The final protein concentration was 3.5 mg/ml. Before mounting, 1 micro-L of a 20mM solution of a small molecule NCGC00015280 (demonstrated as inhibiting the AMA1-RON2 interaction and referenced as B43 in the PDB), was dissolved in DMSO and half diluted with the reservoir buffer, then added to the drop containing the crystals. Cryo-protecting buffer for the PfAMA1-3D7 crystals consisted of 90% of reservoir buffer containing 20% PEG 3350, 0.1M Hepes pH 7 and 10% propanol-2 supplemented with 10% glycerol, and 10% of a 20mM solution of B43.