7OJX

E2 UBE2K covalently linked to donor Ub, acceptor di-Ub, and RING E3 primed for K48-linked Ub chain synthesis

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	DIAMOND BEAMLINE I04-1
Synchrotron site	Diamond
Beamline	I04-1
Temperature [K]	100
Detector technology	PIXEL
Collection date	2020-02-09
Detector	DECTRIS PILATUS 6M-F
Wavelength(s)	0.9119
Spacegroup name	P 31 2 1
Unit cell lengths	85.835, 85.835, 167.354
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	74.340 - 2.400
R-factor	0.1902
Rwork	0.188
R-free	0.23600
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1yla 1ubq 4v3l
Data reduction software	DIALS
Data scaling software	xia2
Phasing software	PHASER
Refinement software	PHENIX (1.17.1_3660)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	74.340	74.370	2.440
High resolution limit [Å]	2.400	6.510	2.400
Rmeas	0.060	0.034	1.102
Rpim	0.020	0.012	0.370
Total number of observations	246998
Number of reflections	28670	1587	1419
<I/σ(I)>	19.1	74.4	0.7
Completeness [%]	100.0	100	99.9
Redundancy	8.6	7.9	8.8

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	7.5	292	Crystals were grown in: 0.2 M sodium citrate, 0.1 M Bis Tris propane 7.5 and 20 % (w/v) PEG 3350. The crystallization drops were set as a 1:1 mixture of the protein complex solution and the precipitant solution.