6RHC

Fragment AZ-003 binding at the TAZpS89/14-3-3 sigma interface

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	PETRA III, DESY BEAMLINE P11
Synchrotron site	PETRA III, DESY
Beamline	P11
Temperature [K]	100
Detector technology	PIXEL
Collection date	2018-10-28
Detector	DECTRIS PILATUS 6M-F
Wavelength(s)	1.033191
Spacegroup name	C 2 2 21
Unit cell lengths	82.025, 112.307, 62.697
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	41.830 - 1.200
R-factor	0.182
Rwork	0.181
R-free	0.19674
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	3mhr
RMSD bond length	0.015
RMSD bond angle	1.912
Data reduction software	DIALS
Data scaling software	Aimless
Phasing software	MOLREP
Refinement software	REFMAC (5.8.0238)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	41.830	1.220
High resolution limit [Å]	1.200	1.200
Rmeas	0.113	0.937
Number of reflections	89802	4345
<I/σ(I)>	11.3	2.1
Completeness [%]	99.3	97.3
Redundancy	12.8	12.8
CC(1/2)	0.997	0.575

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	7.5	277.15	Protein and peptide were mixed at a 1:2 molar stoichiometry with a final protein concentration of 12 mg/mL in crystallization buffer. This was used during hanging-drop crystallization in a 1:1 ratio with 0.1 M HEPES pH 7.5, 0.2 M CaCl2, 5% glycerol, 2 mM BME and 28% PEG400. Crystals were grown within 10 days at 4 C and fragment soaking was performed on crystals of 10 days and older by adding 0.2 uL of a 100 mM stock solution in dimethyl sulfoxide to 2 uL drops containing multiple crystals.