4ESV

A New Twist on the Translocation Mechanism of Helicases from the Structure of DnaB with its Substrates

X-RAY DIFFRACTION

Crystallization

Crystalization Experiments
ID	Method	pH	Temperature	Details
1	VAPOR DIFFUSION, SITTING DROP	6.5	290	4-7% PEG3350, 0.2 M calcium chloride, 0.1 M MES, pH 6.5, VAPOR DIFFUSION, SITTING DROP, temperature 290K

Crystal Properties
Matthews coefficient	Solvent content
3.04	59.59

Crystal Data

Unit Cell
Length ( Å )	Angle ( ˚ )
a = 149.124	α = 90
b = 180.321	β = 90
c = 279.128	γ = 90

Symmetry
Space Group	P 2 2 21

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	ADSC QUANTUM 315			M	SINGLE WAVELENGTH
2	1	x-ray	100	PIXEL	DECTRIS PILATUS 6M		2011-04-04	M	SINGLE WAVELENGTH
3	1	x-ray	100	CCD	ADSC QUANTUM 315		2011-06-17	M	SINGLE WAVELENGTH

Radiation Source
ID #	Source	Type	Wavelength List	Synchrotron Site	Beamline
1	SYNCHROTRON	APS BEAMLINE 24-ID-C	1.11	APS	24-ID-C
2	SYNCHROTRON	NSLS BEAMLINE X25	1.2545,1.2300,1.2549	NSLS	X25
3	SYNCHROTRON	NSLS BEAMLINE X29A	1.11	NSLS	X29A

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
1,2,3	3.2	50	88.3	0.115	6.5	3	124032	124032

Highest Resolution Shell
ID #	Resolution (High)	Resolution (Low)	Percent Possible (All)	Percent Possible (Observed)	Mean I Over Sigma (Observed)	Redundancy	Number Unique Reflections (All)
1,2,3	3.2	3.42	87.6			2.8	9746

Refinement

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 (Observed)	R-Work	R-Free	R-Free Selection Details	Mean Isotropic B
X-RAY DIFFRACTION	SIRAS	THROUGHOUT	3.2	47.92	108863	5442	87.77	0.245	0.242	0.289	RANDOM	140.749

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]
-0.14			0.03		0.11

RMS Deviations
Key	Refinement Restraint Deviation
r_dihedral_angle_2_deg	40.967
r_dihedral_angle_3_deg	21.808
r_dihedral_angle_4_deg	16.874
r_scangle_it	9.194
r_dihedral_angle_1_deg	7.344
r_mcangle_it	5.949
r_scbond_it	5.329
r_mcbond_it	3.266
r_angle_refined_deg	1.506
CHIRAL-CEN RESTRAINTS (A**3)	0.113

RMS Deviations
Key	Refinement Restraint Deviation
r_dihedral_angle_2_deg	40.967
r_dihedral_angle_3_deg	21.808
r_dihedral_angle_4_deg	16.874
r_scangle_it	9.194
r_dihedral_angle_1_deg	7.344
r_mcangle_it	5.949
r_scbond_it	5.329
r_mcbond_it	3.266
r_angle_refined_deg	1.506
CHIRAL-CEN RESTRAINTS (A**3)	0.113
r_bond_refined_d	0.011
r_gen_planes_refined	0.005

Non-Hydrogen Atoms Used in Refinement
Non-Hydrogen Atoms	Number
Protein Atoms	39351
Nucleic Acid Atoms	540
Solvent Atoms	5
Heterogen Atoms	392

Software

Software
Software Name	Purpose
DENZO	data reduction
SCALEPACK	data scaling
SHELX	phasing
SHELX	model building
REFMAC	refinement
PDB_EXTRACT	data extraction
ADSC	data collection
CBASS	data collection
HKL-2000	data reduction
HKL-2000	data scaling
SHELXD	phasing

RCSB PDB Core Operations are funded by the National Science Foundation (DBI-1832184), 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.