3LLU

Crystal structure of the nucleotide-binding domain of Ras-related GTP-binding protein C

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 19-ID
Synchrotron site	APS
Beamline	19-ID
Temperature [K]	100
Detector technology	CCD
Collection date	2009-12-16
Detector	ADSC Q315
Wavelength(s)	0.97934
Spacegroup name	I 4
Unit cell lengths	72.220, 72.220, 72.182
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	22.840 - 1.400
R-factor	0.155
Rwork	0.154
R-free	0.18100
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	2q3f
RMSD bond length	0.015
RMSD bond angle	1.515
Data reduction software	DENZO
Data scaling software	SCALEPACK
Phasing software	PHASER
Refinement software	REFMAC (5.5.0102)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	25.000	25.000	1.420
High resolution limit [Å]	1.400	3.800	1.400
Rmerge	0.055	0.032	0.816
Number of reflections	36466
<I/σ(I)>	12.5
Completeness [%]	100.0	99.3	100
Redundancy	7.3	7.4	7.2

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	7.5	291	2.0M ammonium sulfate, 0.2M sodium chloride, 0.1M HEPES. 1:100 (w/w) papain and GMPPNP were also added. Please note: mass spectral analysis of TEV protease digest prior to crystallization suggested that removal of the His-tag was incomplete even after 2 days. However, without TEV protease treatment no crystals were obtained using otherwise identical crystallization conditions. Also note the presence of papain in the crystallization drop., pH 7.5, vapor diffusion, sitting drop, temperature 291K