4X3G

Crystal structure of SIAH1 SINA domain in complex with a USP19 peptide

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 19-ID
Synchrotron site	APS
Beamline	19-ID
Temperature [K]	100
Detector technology	CCD
Collection date	2014-07-17
Detector	ADSC QUANTUM 315
Wavelength(s)	0.97918
Spacegroup name	P 1 21 1
Unit cell lengths	41.343, 88.092, 59.590
Unit cell angles	90.00, 103.24, 90.00

Refinement procedure

Resolution	48.450 - 2.340
R-factor	0.2233
Rwork	0.222
R-free	0.25120
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	4i7b
RMSD bond length	0.008
RMSD bond angle	0.950
Data reduction software	HKL-3000
Data scaling software	HKL-3000
Phasing software	MOLREP
Refinement software	BUSTER-TNT (BUSTER 2.10.0)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	50.000	50.000	2.370
High resolution limit [Å]	2.330	6.320	2.330
Rmerge	0.089	0.072	0.440
Total number of observations	36140
Number of reflections	15818
<I/σ(I)>	6.2
Completeness [%]	90.9	87.9	58.9
Redundancy	2.3	2.4	1.5

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	9	298	SIAH1 AT 17.6 MG/ML WAS MIXED WITH A TWO-FOLD EXCESS OF USP19 PEPTIDE ON ICE FOR 30 MIN. BEFORE SETTING UP FOR CRYSTALLIZATION. CRYSTALS WERE GROWN AT 298K USING THE SITTING DROP METHOD BY MIXING 0.5 UL PROTEIN:PEPTIDE MIX WITH 0.5 UL WELL SOLUTION CONSISTING OF 20% PEG6000, 0.1 M BICINE PH 9.0. THE CRYSTALS WERE CRYOPROTECTED BY FIRST IMMERSION IN WELL SOLUTION MIXED WITH 15% (FINAL) ETHYLENE GLYCOL, THEN IMMERSION IN N-PARATONE.