6J4U

Structural basis of tubulin detyrosination by vasohibins-SVBP enzyme complex and functional implications

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	SSRF BEAMLINE BL19U1
Synchrotron site	SSRF
Beamline	BL19U1
Temperature [K]	100
Detector technology	PIXEL
Collection date	2018-06-24
Detector	DECTRIS PILATUS3 S 6M
Wavelength(s)	0.9789
Spacegroup name	P 2 21 21
Unit cell lengths	44.512, 71.453, 128.108
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	47.695 - 1.998
R-factor	0.1699
Rwork	0.168
R-free	0.20360
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	6j4o
RMSD bond length	0.007
RMSD bond angle	0.869
Data reduction software	HKL-3000
Data scaling software	HKL-3000
Phasing software	PHASER
Refinement software	PHENIX ((1.12_2829: ???))

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	50.000	2.070
High resolution limit [Å]	1.998	2.000
Rmerge	0.073	1.000
Rpim	0.023	0.358
Number of reflections	28511	2759
<I/σ(I)>	24.6	1.2
Completeness [%]	99.9	99.5
Redundancy	12.4	9.5
CC(1/2)		0.738

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	EVAPORATION	5	293	by introducing three mutations E71S/A72H/K79M into V1c allowed us to solve the V1c-SVBP to high resolution. Well diffracting crystals of the mutant V1c-SVBP complex were obtained in 1.0 M lithium chloride, 0.1 M citric acid, pH 5.0, 20% PEG 6000.