5HI7

Co-crystal structure of human SMYD3 with an aza-SAH compound

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 21-ID-D
Synchrotron site	APS
Beamline	21-ID-D
Temperature [K]	295
Detector technology	CCD
Collection date	2013-01-13
Detector	MARMOSAIC 300 mm CCD
Wavelength(s)	1.07820
Spacegroup name	P 21 21 21
Unit cell lengths	60.470, 66.401, 104.851
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	44.709 - 2.150
R-factor	0.1999
Rwork	0.198
R-free	0.22570
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.003
RMSD bond angle	0.626
Data reduction software	HKL-2000
Data scaling software	HKL-2000
Phasing software	PHENIX
Refinement software	PHENIX (dev_1801)

Data quality characteristics

	Overall
Low resolution limit [Å]	50.000
High resolution limit [Å]	2.100
Number of reflections	46288
<I/σ(I)>	11.4
Completeness [%]	99.2
Redundancy	2.2

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP		295	SMYD3 crystallization and soaking of GSK2807: SMYD3 (1-428) expressed in baculovirus and with SAH in the SAM-binding site was crystallized in sitting drops at 22C using 1uL fresh protein at 8.9mg/mL with the addition of 1uL mother liquor (200mM MgOAc Tetrahydrate, 20% PEG 3350). Crystals grew to a large size but varied in quantity when set up in replicates. A seed stock was formed from these crystals and added at 20% to the mother liquor solution prior to plate setup. Crystals of large size were then created with 1.3uL protein and 1.3uL of mother liquor containing seeds in a sitting drop MRC-2 plate (Hampton Research). Crystal nucleation occurred overnight and reached maximum size in 4 days