8CCZ

Crystal structure of human Sirt3 in complex with an inhibiting HIV1 Tat-37-59 peptide

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	BESSY BEAMLINE 14.1
Synchrotron site	BESSY
Beamline	14.1
Temperature [K]	100
Detector technology	PIXEL
Collection date	2018-06-13
Detector	DECTRIS PILATUS 6M
Wavelength(s)	0.9184
Spacegroup name	P 1 21 1
Unit cell lengths	54.545, 78.124, 76.575
Unit cell angles	90.00, 96.09, 90.00

Refinement procedure

Resolution	54.240 - 1.950
R-factor	0.2293
Rwork	0.227
R-free	0.27630
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	4bvb
RMSD bond length	0.002
RMSD bond angle	0.511
Data reduction software	DIALS
Data scaling software	Aimless
Phasing software	PHASER
Refinement software	PHENIX (1.17.1_3660)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	54.240	2.020
High resolution limit [Å]	1.950	1.950
Number of reflections	46174	4615
<I/σ(I)>	11.97	0.55
Completeness [%]	98.8	98.88
Redundancy	5.5	5.5
CC(1/2)	0.949	0.445

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP		293.15	10 mg/ml human Sirt3-(118-399) in 20 mM Tris/HCl, pH 8.0, 150 mM NaCl, 5% (v/v) glycerol, 1 mM TCEP were incubated with 2 mM Tat-37-59 in 10% (v/v) DMSO for 60 min at 293.15 K. The complex was crystallized using the sitting-drop vapor-diffusion method at 293.15 K with 100 mM CHES, pH 9.0, 20% (w/v) PEG 8000 as reservoir solution.