4EV0

Crystal Structure of Thermus thermophilus Catabolite Activator Protein

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	NSLS BEAMLINE X29A
Synchrotron site	NSLS
Beamline	X29A
Temperature [K]	100
Detector technology	CCD
Collection date	2010-10-07
Detector	ADSC QUANTUM 315
Wavelength(s)	1.075
Spacegroup name	P 21 21 21
Unit cell lengths	46.250, 94.842, 121.439
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	44.173 - 2.402
R-factor	0.2028
Rwork	0.200
R-free	0.25550
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	Nterminal domain: composite model of pruned fragments from PDB entries 2pqq 3d0s 3h3u representing residues 21-102; C-terminal domain: pruned fragment from PDB entry 2zcw
RMSD bond length	0.008
RMSD bond angle	1.165
Data reduction software	DENZO
Data scaling software	SCALEPACK
Phasing software	PHASER
Refinement software	PHENIX (1.7.3_928)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	100.000	100.000	2.440
High resolution limit [Å]	2.400	6.510	2.400
Rmerge	0.080	0.045	0.565
Number of reflections	20242
<I/σ(I)>	13.9
Completeness [%]	93.9	99.3	50.6
Redundancy	24.5	25.2	6.5

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	7	293	Screening for crystallization used the Gryphon liquid handling system (Art Robbins Instruments). Crystals were grown using sitting drops (200 nL protein + 200 nL crystallization reagent) with 60 UL reservoirs of crystallization reagent in a 96-well high-throughput screen. Plate-shaped crystals were obtained at 20 degree C using Hampton Research NATRIX HT #38: 0.2 M ammonium acetate, 0.15 M magnesium acetate tetrahydrate, 5% (w/v) polyethylene glycol 4000, and 0.05 M HEPES sodium, pH 7.0, VAPOR DIFFUSION, SITTING DROP, temperature 293K