7JIG

HRAS A59T GppNHp crystal 2

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	ROTATING ANODE
Source details	RIGAKU MICROMAX-007 HF
Temperature [K]	173.15
Detector technology	IMAGE PLATE
Collection date	2016-03-04
Detector	RIGAKU RAXIS IV++
Wavelength(s)	1.54178
Spacegroup name	P 32 2 1
Unit cell lengths	38.964, 38.964, 159.048
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	33.744 - 2.322
R-factor	0.1868
Rwork	0.177
R-free	0.26350
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1ctq
RMSD bond length	0.007
RMSD bond angle	0.916
Data reduction software	HKL-3000
Data scaling software	HKL-3000
Phasing software	PHENIX (1.11.1-2575)
Refinement software	PHENIX (1.11.1_2575)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	50.000	50.000	2.360
High resolution limit [Å]	2.320	6.290	2.320
Rmerge	0.076	0.043	0.420
Rmeas	0.082	0.047	0.516
Rpim	0.031	0.018	0.292
Number of reflections	6642	404	231
<I/σ(I)>	9.5
Completeness [%]	96.7	99.8	75.5
Redundancy	6.5	6.2	2.2
CC(1/2)		0.998	0.765

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	7.5	291.15	Starting concentration of HRAS A59T GppNHp was 12.1mg/mL in buffer containing 20mM HEPES, 50mM NaCl, 20mM MgCl2 and 1mM DTT at pH 7.5. Used a 24-well plate sealed with Vaseline and total well volumes of 0.402mL. Hanging drops were 0.001mL mother liquor to 0.001mL protein. Mother liquor contained 3.4mM NaCl, 1mM MgCl2, 15.7mM HEPES (pH7.5), 2.5mM DTT, 10mM Mg(OAc)2, and 44.8% PEG 400.