4DVG

Crystal structure of E. histolytica Formin1 bound to EhRho1-GTPgammaS

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 23-ID-B
Synchrotron site	APS
Beamline	23-ID-B
Temperature [K]	100
Detector technology	IMAGE PLATE
Collection date	2011-11-22
Detector	MAR scanner 300 mm plate
Wavelength(s)	0.9795
Spacegroup name	P 61
Unit cell lengths	138.564, 138.564, 57.765
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	35.673 - 2.604
R-factor	0.2188
Rwork	0.215
R-free	0.25120
Structure solution method	SAD
RMSD bond length	0.010
RMSD bond angle	1.233
Data reduction software	HKL-2000
Data scaling software	HKL-2000
Phasing software	PHENIX (AutoSol)
Refinement software	PHENIX ((phenix.refine: 1.7.1_743))

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	40.000	2.630
High resolution limit [Å]	2.600	2.600
Rmerge	0.085	0.580
Number of reflections	19500
<I/σ(I)>		2.1
Completeness [%]	98.4
Redundancy	9.6

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	8.5	293	EhRho1-GTPgammaS at 5 mg/mL and and EhFormin1 at 10 mg/mL were mixed in crystallization buffer (50 mM Tris pH 8.0, 250 mM NaCl, 2.5% (v/v) glycerol, 5 mM DTT, 50 microM GTPgammaS, 1 mM magnesium chloride) and allowed to form a complex for 30 minutes at room temperature. The protein solution was then mixed 1:1 and equilibrated against crystallization solution (18% PEG 3350, 100 mM Tris pH 8.5, 200 mM magnesium chloride)., VAPOR DIFFUSION, HANGING DROP, temperature 293K