5LBX

Structure of the T175V Etr1p mutant in the trigonal form P312 in complex with NADP and crotonyl-CoA

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	SLS BEAMLINE X10SA
Synchrotron site	SLS
Beamline	X10SA
Temperature [K]	100
Detector technology	PIXEL
Collection date	2015-12-01
Detector	DECTRIS PILATUS 6M-F
Wavelength(s)	1.73913
Spacegroup name	P 31 2 1
Unit cell lengths	93.498, 93.498, 231.203
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	46.749 - 2.500
R-factor	0.1835
Rwork	0.182
R-free	0.21240
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	4w99
RMSD bond length	0.007
RMSD bond angle	1.002
Data reduction software	XDS
Data scaling software	SCALA
Phasing software	PHASER
Refinement software	PHENIX ((1.10.1_2155: ???))

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	47.040	2.640
High resolution limit [Å]	2.500	2.500
Rmerge	0.124	1.511
Number of reflections	41476
<I/σ(I)>	12.2	1.5
Completeness [%]	100.0	100
Redundancy	8.6	7.7
CC(1/2)	0.997

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	6.4	298	Etr1p was crystallized at 15 mg/mL with 5 mM NADP and 5 mM of crotonyl-CoA in the gel filtration buffer (100 mM NaCl, 20 mM Tris-HCl pH 7.9). Crystals appeared after several days in 2.0 M (NH4)2SO4, 100 mM ADA/NaOH pH 6.4 at room temperature. The crystals were soaked in a solution containing 30% (v/v) glycerol, 2.0 M (NH4)2SO4, 100 mM ADA pH 6.4 prior to freezing in liquid nitrogen to prevent water-crystal formation.