Experiment: 8BWG

X-RAY DIFFRACTION

Initial Refinement Model(s)
Type	Source	Accession Code	Details
experimental model	PDB	1WQ1

Crystalization Experiments
ID	Method	pH	Temperature	Details
1	VAPOR DIFFUSION, SITTING DROP		293	Using a commercial crystal screen (HAMPTON RESEARCH, HR2-130) yielded a hit for monophosphorylated HRas under sitting drop conditions (drop size 600 nL) with a 1:1 ratio of protein solution (phospho-HRas 0.4 mM, RasGAP 0.4 mM, Na-HEPES 20 mM pH = 8.0, MgCl2 5 mM, NaF 20 mM) and precipitant (Na-citrate 100 mM pH = 5.6, Li2SO4 1.0 M, CaCl2 200 mM). After three rounds of microseeding well-formed single crystals were obtained using 2.0 uL sitting drops and a 1:1 ratio of protein buffer (HRas 400 uM, RasGAP 400 uM, MgCl2 5 mM, Na-HEPES 20 mM pH = 8.0, NaF 20 mM) and precipitant (Na-Citrate 100 mM pH = 5.6, Li2SO4 800 mM, CaCl2 200 mM). These were harvested using cryoprotectant (80% precipitant, 20% glycerol (v/v)) and sent for data collection.

Crystal Properties
Matthews coefficient	Solvent content
2.61	52.9

Symmetry
Space Group	H 3 2

Diffraction Experiment
ID #	Crystal ID	Scattering Type	Data Collection Temperature	Detector	Detector Type	Details	Collection Date	Monochromator	Protocol
1	1	x-ray	100	PIXEL	DECTRIS EIGER2 XE 16M		2021-01-29	M	SINGLE WAVELENGTH

Radiation Source
ID #	Source	Type	Wavelength List	Synchrotron Site	Beamline
1	SYNCHROTRON	DIAMOND BEAMLINE I03	0.9763	Diamond	I03

Overall
ID #	Resolution (High)	Resolution (Low)	Percent Possible (Observed)	R Merge I (Observed)	Rrim I (All)	Rpim I (All)	CC (Half)	Net I Over Average Sigma (I)	Redundancy	Number Reflections (All)	Number Reflections (Observed)	Observed Criterion Sigma (F)	Observed Criterion Sigma (I)	B (Isotropic) From Wilson Plot
1	1.32	47.925	99.2	0.041	0.043	0.014	1	28.6	17.6		45988

Highest Resolution Shell
ID #	Resolution (High)	Resolution (Low)	Percent Possible (All)	Percent Possible (Observed)	R Merge I (Observed)	Rrim I (All)	Rpim I (All)	CC (Half)	Mean I Over Sigma (Observed)	Redundancy	Number Unique Reflections (All)
1	1.32	1.34			1.34	1.535	0.723	0.577		7.8

Statistics
Diffraction ID	Structure Solution Method	Cross Validation method	Resolution (High)	Resolution (Low)	Number Reflections (Observed)	Number Reflections (R-Free)	Percent Reflections (Observed)	R-Factor (All)	R-Work	R-Free	Mean Isotropic B
X-RAY DIFFRACTION	MOLECULAR REPLACEMENT	FREE R-VALUE	1.32	47.925	45983	2232	99.176	0.137	0.1354	0.1739	22.946

Temperature Factor Modeling
Anisotropic B[1][1]	Anisotropic B[1][2]	Anisotropic B[1][3]	Anisotropic B[2][2]	Anisotropic B[2][3]	Anisotropic B[3][3]
-0.796	-0.398		-0.796		2.583

RMS Deviations
Key	Refinement Restraint Deviation
r_dihedral_angle_6_deg	16.648
r_rigid_bond_restr	15.682
r_dihedral_angle_3_deg	14.448
r_dihedral_angle_2_deg	8.675
r_scbond_other	7.62
r_scbond_it	7.604
r_scangle_it	7.202
r_scangle_other	7.192
r_dihedral_angle_1_deg	6.465
r_lrange_it	5.369

RMS Deviations
Key	Refinement Restraint Deviation
r_dihedral_angle_6_deg	16.648
r_rigid_bond_restr	15.682
r_dihedral_angle_3_deg	14.448
r_dihedral_angle_2_deg	8.675
r_scbond_other	7.62
r_scbond_it	7.604
r_scangle_it	7.202
r_scangle_other	7.192
r_dihedral_angle_1_deg	6.465
r_lrange_it	5.369
r_lrange_other	5.079
r_mcangle_it	3.612
r_mcangle_other	3.61
r_mcbond_it	3.283
r_mcbond_other	3.171
r_angle_refined_deg	1.607
r_angle_other_deg	0.592
r_nbd_refined	0.266
r_xyhbond_nbd_refined	0.222
r_symmetry_nbd_other	0.201
r_nbtor_refined	0.18
r_symmetry_xyhbond_nbd_refined	0.174
r_symmetry_nbd_refined	0.163
r_nbd_other	0.147
r_chiral_restr	0.089
r_symmetry_nbtor_other	0.083
r_bond_refined_d	0.01
r_gen_planes_refined	0.009
r_gen_planes_other	0.002
r_bond_other_d	0.001