8CCW

Crystal structure of human Sirt3 in complex with an acetylated HIV1 Tat-46-54 substrate 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	2018-07-12
Detector	DECTRIS PILATUS 6M
Wavelength(s)	0.9184
Spacegroup name	C 2 2 21
Unit cell lengths	78.721, 127.760, 77.576
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	39.360 - 1.650
R-factor	0.1813
Rwork	0.181
R-free	0.20890
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	4fvt
RMSD bond length	0.017
RMSD bond angle	1.382
Data reduction software	XDS
Data scaling software	XDS
Phasing software	PHASER
Refinement software	PHENIX (1.17.1_3660)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	39.360	1.709
High resolution limit [Å]	1.650	1.650
Number of reflections	47332	4698
<I/σ(I)>	13.93	0.89
Completeness [%]	99.9	99.94
Redundancy	11.2	11.2
CC(1/2)	0.999	0.469

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 ac-Tat-46-54 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 MES, pH 6.0, 30% (w/v) PEG 200, 5% (w/v) PEG 3000 as reservoir solution.