6M8Q

Cleavage and Polyadenylation Specificity Factor Subunit 3 (CPSF3) in complex with NVP-LTM531

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 17-ID
Synchrotron site	APS
Beamline	17-ID
Temperature [K]	100
Detector technology	PIXEL
Collection date	2014-03-19
Detector	DECTRIS PILATUS3 6M
Wavelength(s)	1
Spacegroup name	P 43 21 2
Unit cell lengths	106.222, 106.222, 206.035
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	73.950 - 2.490
R-factor	0.1654
Rwork	0.163
R-free	0.22080
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	2i7t
RMSD bond length	0.010
RMSD bond angle	1.180
Data reduction software	DENZO
Data scaling software	SCALEPACK
Phasing software	PHASER
Refinement software	BUSTER (2.11.4)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	94.410	2.500
High resolution limit [Å]	2.490	2.490
Rmerge	0.138	1.272
Number of reflections	42063
<I/σ(I)>	12.6	2.2
Completeness [%]	99.7	100
Redundancy	7.1	7.1

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	7	294	Reservoir solution: 100 mM Tris HCl pH 7.0; 0.4 M NaH2PO4; 1.466 K2HPO; 0.2 M NaCl 10 mg/mL Protein solution: CPSF3 protein in; 20 mM Hepes pH 7.7; 150 mM NaCl; 5 % glycerol; 1 mM DTT; 0.5 mM NVP-LTM531 Co-crystallization Protocol: 50 nL of protein mixed with 50 nL of reservoir solution. Crystals grew within 1 day with tetragonal bipyramidal habitus and reached a maximum size of 50 micrometers after 3 days. Cryo-protection Protocol: supplemented reservoir solution with 10 and 20% glycerol, respectively, and subsequently added both solution within 5 min. Crystals incubate for another 5 min before harvesting.