4XFK

Crystal structure of Leucine-, Isoleucine-, Valine-, Threonine-, and Alanine-binding protein from Brucella ovis

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 21-ID-F
Synchrotron site	APS
Beamline	21-ID-F
Temperature [K]	100
Detector technology	CCD
Collection date	2010-11-12
Detector	RAYONIX MX-225
Wavelength(s)	0.97872
Spacegroup name	P 1 21 1
Unit cell lengths	62.120, 46.390, 62.400
Unit cell angles	90.00, 101.62, 90.00

Refinement procedure

Resolution	30.424 - 1.300
R-factor	0.1379
Rwork	0.137
R-free	0.16110
Structure solution method	SAD
Starting model (for MR)	structure initially solved via Iodide SAD in oP crystal form this model was used for Molecular Replacement into mP crystal form
RMSD bond length	0.006
RMSD bond angle	1.098
Data reduction software	XDS
Data scaling software	XSCALE
Phasing software	PHASER
Refinement software	PHENIX

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	50.000		1.330
High resolution limit [Å]	1.300	5.810	1.300
Rmerge	0.046	0.018	0.517
Rmeas	0.054	0.021	0.605
Total number of observations	314986
Number of reflections	83917	988	6070
<I/σ(I)>	17.96	53.65	2.49
Completeness [%]	97.9	96.5	96.1
Redundancy	3.8		3.7

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	9.5	290	Microlytics MCSG1 screen, a2: 100mM CHES/NaOH, pH 9.5, 30% PEG 3000; BrovA.17370.a.B2.PS02137 at 25mg/ml; cryo: Al's oil; tray 257610a2, puck wln2-3; crystal #2 was used for phasing
2	VAPOR DIFFUSION, SITTING DROP	7.5	290	Microlytics MCSG1 screen, c10: 1M LiCl, 100mM Na-acetate, 30% PEG 6000; the crystal was incubated in two cryo/phasing solution for 30sec each: 4.5ul reservoir + 0.5ul of 2.5M NaI in ethylene glycol, and 4ul reservoir + 1ul of 2.5M NaI in ethylene glycol, final 500mM NaI and 20% ethylene glycol; this is a different crystal form: P212121 with a=60.7AA, b=67.26AA, c=94.5AA; this crystal was used for phasing; tray 257610c10, puck ilg3-2