4LHZ

Crystal structure of GTP-bound Rab8:Rabin8

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	SLS BEAMLINE X10SA
Synchrotron site	SLS
Beamline	X10SA
Temperature [K]	100
Detector technology	CCD
Collection date	2009-05-01
Detector	MARMOSAIC 225 mm CCD
Wavelength(s)	1.0000
Spacegroup name	C 2 2 21
Unit cell lengths	81.337, 165.561, 167.263
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	19.990 - 3.200
R-factor	0.27918
Rwork	0.278
R-free	0.30877
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1yzq
RMSD bond length	0.016
RMSD bond angle	1.639
Data reduction software	MOSFLM
Data scaling software	SCALEPACK
Phasing software	PHASES
Refinement software	REFMAC (5.2.0019)

Data quality characteristics

	Overall
Low resolution limit [Å]	20.000
High resolution limit [Å]	3.200
Number of reflections	18932
Completeness [%]	99.4

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	6.6	293	Nucleotide-free Rab8a1 184:Rabin8157-232 was crystallized in 18% (w/v) PEG3350, 0.1 M Li2SO4, 0.1 M MES pH 6.6. Before data collection, the complex crystal was protected with cryo solution (30% (w/v) PEG3350, 0.1 M Li2SO4, 0.1 M MES pH 6.6). In order to produce nucleotide-bound forms of Rab8:Rabin8 complexes, the nucleotide-free Rab8:Rabin8 crystals were soaked with cryo solution containing 30 % (w/v) PEG3350, 0.1 M Li2SO4, 0.1 M MES pH 6.6 and 1 mM respective nucleotide GDP/GTP for one hours at 278 K, VAPOR DIFFUSION, HANGING DROP, temperature 293K