7T1D

Human SIRT2 in complex with small molecule 359

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ESRF BEAMLINE MASSIF-1
Synchrotron site	ESRF
Beamline	MASSIF-1
Temperature [K]	100
Detector technology	PIXEL
Collection date	2020-09-01
Detector	DECTRIS PILATUS 6M-F
Wavelength(s)	1.0332
Spacegroup name	P 1 21 1
Unit cell lengths	36.775, 55.980, 139.576
Unit cell angles	90.00, 94.46, 90.00

Refinement procedure

Resolution	35.002 - 1.750
Rwork	0.163
R-free	0.21280
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	CRELUX reference structure of SIRT2
RMSD bond length	0.010
RMSD bond angle	1.574
Data reduction software	XDS
Data scaling software	Aimless (0.7.4)
Phasing software	PHASER
Refinement software	REFMAC (5.8.0267)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	139.150	139.150	1.780
High resolution limit [Å]	1.750	9.090	1.750
Rmerge	0.075	0.040	0.612
Rmeas	0.089	0.048	0.727
Rpim	0.047	0.025	0.388
Total number of observations	194963	1502	10269
Number of reflections	56312	443	3005
<I/σ(I)>	9.5	26.7	1.9
Completeness [%]	98.6	99.6	98.9
Redundancy	3.5	3.4	3.4
CC(1/2)	0.996	0.992	0.664

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	8	293.15	Crystals of SIRT2 in complex with FLS-359 were obtained using hanging-drop vapour diffusion setups. SIRT2 at a concentration of 21.9 mg/ml (50 mM Hepes-NaOH, 150 mM NaCl, pH 8.0) was preincubated with 3.6 mM (5.7-fold molar excess) of FLS-359 (100 mM in DMSO) for 1 h. 1 ul of the protein solution was then mixed with 2 ul of reservoir solution (0.1 M Hepes-NaOH pH 6.6, 0.3 M Li2SO4, 21 % (w/v) PEG 3350) and streak seeded before being equilibrated at 20C over 0.2 ml of reservoir solution. Well diffracting crystals grew as thick aggregates of thin plates and were mounted within 19 days