8BR0

ExoY Nucleotidyl Cyclase domain from Vibrio nigripulchritudo MARTX toxin (residue Q3455 to L3863) in complex with 3'deoxyCTP and two manganese cations bound to Latrunculin-B-ADP-Mn-actin


X-RAY DIFFRACTION

Crystallization

Crystalization Experiments
IDMethodpHTemperatureDetails
1VAPOR DIFFUSION, HANGING DROP29110mg/mL binary complex in the presence of 15.2 mM 3primedCTP1.6 mM ADP, 20 mM MgCl2, 0.2 mM Latrunculin B, 23 mM KCl, 70 mM LiCl, 8 mM HEPES pH 8.5, 4 mM TCEP and mixed with 17 % PEG4000, 17 % Glycerol, 0.01 M Li2SO4, 0.1 M Tris pH 8.5, 5 mM MgCl2, 15 mM MnCl2, 1% 1-Butyl-2,3-dimethylimidazolium tetrafluoroborate (ionic liquid 18 from the Ionic Liquid Screen (Hampton Research)) in a 1:1.2 v/v hanging drop
Crystal Properties
Matthews coefficientSolvent content
2.6353.18

Crystal Data

Unit Cell
Length ( Å )Angle ( ˚ )
a = 74.469α = 90
b = 132.503β = 110.4
c = 96.217γ = 90
Symmetry
Space GroupP 1 21 1

Diffraction

Diffraction Experiment
ID #Crystal IDScattering TypeData Collection TemperatureDetectorDetector TypeDetailsCollection DateMonochromatorProtocol
11x-ray100PIXELDECTRIS EIGER X 16MBimorph Kirkpatrick-Baez mirrors2022-04-22MSINGLE WAVELENGTH
Radiation Source
ID #SourceTypeWavelength ListSynchrotron SiteBeamline
1SYNCHROTRONSOLEIL BEAMLINE PROXIMA 10.97856SOLEILPROXIMA 1

Data Collection

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)RedundancyNumber Reflections (All)Number Reflections (Observed)Observed Criterion Sigma (F)Observed Criterion Sigma (I)B (Isotropic) From Wilson Plot
12.21848.05292.20.0980.1050.0360.99811.88.46205964.232
Highest Resolution Shell
ID #Resolution (High)Resolution (Low)Percent Possible (All)Percent Possible (Observed)Rpim I (All)CC (Half)Mean I Over Sigma (Observed)RedundancyNumber Unique Reflections (All)
12.2182.43749.50.4710.591.58.7

Refinement

Statistics
Diffraction IDStructure Solution MethodCross Validation methodStarting modelResolution (High)Resolution (Low)Number Reflections (Observed)Number Reflections (R-Free)Percent Reflections (Observed)R-Factor (Observed)R-WorkR-FreeR-Free Selection DetailsMean Isotropic B
X-RAY DIFFRACTIONMOLECULAR REPLACEMENTTHROUGHOUT1PAV2.21848.0562059297471.80.21020.20850.2428RANDOM72.02
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]
2.88890.2055-1.8311-1.0578
RMS Deviations
KeyRefinement Restraint Deviation
t_other_torsion18.29
t_omega_torsion3.03
t_angle_deg0.97
t_bond_d0.008
t_dihedral_angle_d
t_trig_c_planes
t_gen_planes
t_it
t_nbd
t_improper_torsion
RMS Deviations
KeyRefinement Restraint Deviation
t_other_torsion18.29
t_omega_torsion3.03
t_angle_deg0.97
t_bond_d0.008
t_dihedral_angle_d
t_trig_c_planes
t_gen_planes
t_it
t_nbd
t_improper_torsion
t_pseud_angle
t_chiral_improper_torsion
t_sum_occupancies
t_utility_distance
t_utility_angle
t_utility_torsion
t_ideal_dist_contact
Non-Hydrogen Atoms Used in Refinement
Non-Hydrogen AtomsNumber
Protein Atoms11911
Nucleic Acid Atoms
Solvent Atoms541
Heterogen Atoms170

Software

Software
Software NamePurpose
BUSTERrefinement
PDB_EXTRACTdata extraction
AutoProcessdata reduction
Aimlessdata scaling
PHASERphasing