5BPT

Atomic-resolution structures of the APC/C subunits Apc4 and the Apc5 N-terminal domain

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	DIAMOND BEAMLINE I24
Synchrotron site	Diamond
Beamline	I24
Temperature [K]	100
Detector technology	CCD
Collection date	2013-05-01
Detector	ADSC QUANTUM 4
Wavelength(s)	0.969
Spacegroup name	P 41
Unit cell lengths	90.469, 90.469, 115.593
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	48.710 - 3.200
R-factor	0.2254
Rwork	0.223
R-free	0.26530
Structure solution method	MAD
RMSD bond length	0.008
RMSD bond angle	1.070
Data reduction software	XDS
Data scaling software	SCALA
Phasing software	SHARP
Refinement software	BUSTER-TNT (2.10.1)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	29.300	3.470
High resolution limit [Å]	3.200	3.200
Rmerge	0.200	0.800
Number of reflections	15477
<I/σ(I)>	13.3	1.6
Completeness [%]	99.8	98.5
Redundancy	7.5	7.7

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	8	293	Apc4 was concentrated to 3.5 mg/ml in a buffer of 20 mM Hepes (pH 8.0), 200 mM NaCl and 2 mM DTT. Initial crystal was obtained by vapour diffusion in setting drop in a buffer containing 0.1 M sodium citrate 5.0, 8% (w/v) PEG 8K. By seeding with the initial crystals, large crystals were grown in a buffer containing 0.1 M sodium citrate 5.0, 3% (w/v) PEG 8K, 250 mM magnesium acetate, 10 mM Tris.HCl (pH 8.5), 200 mM NDSB-211, 4% (v/v) ethylene glycol and 2 mM DTT.