6AM0

Crystal structure of K. lactis Edc1-Dcp1-Dcp2-Edc3 decapping complex with synthetic cap substrate analog

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ALS BEAMLINE 8.3.1
Synchrotron site	ALS
Beamline	8.3.1
Temperature [K]	100
Detector technology	PIXEL
Collection date	2017-05-18
Detector	DECTRIS PILATUS3 S 6M
Wavelength(s)	1.11583
Spacegroup name	C 1 2 1
Unit cell lengths	174.260, 83.250, 104.300
Unit cell angles	90.00, 93.62, 90.00

Refinement procedure

Resolution	45.960 - 2.840
R-factor	0.227
Rwork	0.225
R-free	0.24800
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	5lop
RMSD bond length	0.003
RMSD bond angle	0.721
Data reduction software	XDS (Nov 1, 2016)
Data scaling software	XSCALE (Nov 1, 2016)
Phasing software	PHASER (2.5.7)
Refinement software	PHENIX ((1.10.1_2155: ???))

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	50.000	3.000
High resolution limit [Å]	2.840	2.840
Rmerge	0.058	0.780
Number of reflections	68126
<I/σ(I)>	9.82	1.3
Completeness [%]	98.7	99.1
Redundancy	2.9	2.8

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP		277	Protein solution: 4mg/mL Kl Dcp1/Dcp2/Edc3, 1mM Edc1 peptide, 6mM substrate analog, 10mM magnsium chloride. Well solution: 0.22M magnesium chloride, 0.03M EDTA, 8% PEG 8000. Procedure: 250nL protein solution + 250nL well solution + 50nL 1:10000 seed stock. Set up at RT, grow crystals at 4C.