2ZQB

Crystal structure of a psychrotrophic RNaseHI variant with sextuple thermostabilizing mutations

X-RAY DIFFRACTION

Crystallization

Crystalization Experiments
ID	Method	pH	Temperature	Details
1	VAPOR DIFFUSION, SITTING DROP	6.5	277	30% PEG, MME 5000, 0.1M MES, 0.2M ammonium sulfate, pH 6.5, VAPOR DIFFUSION, SITTING DROP, temperature 277K

Crystal Properties
Matthews coefficient	Solvent content
2.25	45.35

Crystal Data

Unit Cell
Length ( Å )	Angle ( ˚ )
a = 68.239	α = 90
b = 68.239	β = 90
c = 272.816	γ = 90

Symmetry
Space Group	P 41 21 2

Diffraction

Diffraction Experiment
ID #	Crystal ID	Scattering Type	Data Collection Temperature	Detector	Detector Type	Details	Collection Date	Monochromator	Protocol
1	1	x-ray	100	CCD	Bruker DIP-6040	mirrors	2008-07-02	M	SINGLE WAVELENGTH

Radiation Source
ID #	Source	Type	Wavelength List	Synchrotron Site	Beamline
1	SYNCHROTRON	SPRING-8 BEAMLINE BL44XU	1.0	SPring-8	BL44XU

Data Collection

Overall
ID #	Resolution (High)	Resolution (Low)	Percent Possible (Observed)	R Merge I (Observed)	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	2.49	50	100	0.146	28.4	13.9		23782

Highest Resolution Shell
ID #	Resolution (High)	Resolution (Low)	Percent Possible (All)	Percent Possible (Observed)	R Merge I (Observed)	Mean I Over Sigma (Observed)	Redundancy	Number Unique Reflections (All)
	2.49	2.59	100		0.525	6.39	14.2	2299

Refinement

Statistics
Diffraction ID	Structure Solution Method	Cross Validation method	Starting model	Resolution (High)	Resolution (Low)	Number Reflections (Observed)	Number Reflections (R-Free)	Percent Reflections (Observed)	R-Factor (Observed)	R-Work	R-Free	R-Free Selection Details	Mean Isotropic B
X-RAY DIFFRACTION	MOLECULAR REPLACEMENT	THROUGHOUT	PDB ENTRY 2E4L	2.49	47.51	22450	1212	99.7	0.19583	0.19312	0.2472	RANDOM	28.584

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]
0.09			0.09		-0.19

RMS Deviations
Key	Refinement Restraint Deviation
r_dihedral_angle_2_deg	38.254
r_dihedral_angle_4_deg	21.443
r_dihedral_angle_3_deg	20.456
r_dihedral_angle_1_deg	8.541
r_scangle_it	5.354
r_scbond_it	3.551
r_mcangle_it	2.533
r_angle_refined_deg	2.51
r_mcbond_it	1.601
r_symmetry_hbond_refined	0.407

RMS Deviations
Key	Refinement Restraint Deviation
r_dihedral_angle_2_deg	38.254
r_dihedral_angle_4_deg	21.443
r_dihedral_angle_3_deg	20.456
r_dihedral_angle_1_deg	8.541
r_scangle_it	5.354
r_scbond_it	3.551
r_mcangle_it	2.533
r_angle_refined_deg	2.51
r_mcbond_it	1.601
r_symmetry_hbond_refined	0.407
r_symmetry_vdw_refined	0.336
r_nbtor_refined	0.322
r_nbd_refined	0.265
r_xyhbond_nbd_refined	0.19
r_chiral_restr	0.178
r_bond_refined_d	0.03
r_gen_planes_refined	0.01

Non-Hydrogen Atoms Used in Refinement
Non-Hydrogen Atoms	Number
Protein Atoms	4702
Nucleic Acid Atoms
Solvent Atoms	224
Heterogen Atoms	15

Software

Software
Software Name	Purpose
REFMAC	refinement
HKL-2000	data collection
HKL-2000	data reduction
HKL-2000	data scaling
MOLREP	phasing

RCSB PDB Core Operations are funded by the U.S. National Science Foundation (DBI-2321666), the US Department of Energy (DE-SC0019749), and the National Cancer Institute, National Institute of Allergy and Infectious Diseases, and National Institute of General Medical Sciences of the National Institutes of Health under grant R01GM133198.