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

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	SOLEIL BEAMLINE PROXIMA 1
Synchrotron site	SOLEIL
Beamline	PROXIMA 1
Temperature [K]	100
Detector technology	PIXEL
Collection date	2022-04-22
Detector	DECTRIS EIGER X 16M
Wavelength(s)	0.97856
Spacegroup name	P 1 21 1
Unit cell lengths	74.469, 132.503, 96.217
Unit cell angles	90.00, 110.40, 90.00

Refinement procedure

Resolution	48.050 - 2.218
R-factor	0.2102
Rwork	0.208
R-free	0.24280
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1pav
RMSD bond length	0.008
RMSD bond angle	0.970
Data reduction software	AutoProcess (1.0.5)
Data scaling software	Aimless (version 0.7.7)
Phasing software	PHASER (v1.0)
Refinement software	BUSTER (2.10.4 (3-FEB-2022))

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	48.052	2.437
High resolution limit [Å]	2.218	2.218
Rmerge	0.098
Rmeas	0.105
Rpim	0.036	0.471
Number of reflections	62059	2919
<I/σ(I)>	11.8	1.5
Completeness [%]	92.2	49.5
Redundancy	8.4	8.7
CC(1/2)	0.998	0.590

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP		291	10mg/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