2R0G

Chromopyrrolic acid-soaked RebC with bound 7-carboxy-K252c

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 24-ID-C
Synchrotron site	APS
Beamline	24-ID-C
Temperature [K]	141.4
Detector technology	CCD
Collection date	2007-03-21
Detector	ADSC QUANTUM 315
Wavelength(s)	0.9795
Spacegroup name	P 1 21 1
Unit cell lengths	64.892, 78.437, 123.625
Unit cell angles	90.00, 99.61, 90.00

Refinement procedure

Resolution	50.000 - 2.370
Rwork	0.205
R-free	0.25530
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	2r0c
RMSD bond length	0.007
RMSD bond angle	1.304
Data reduction software	DENZO
Data scaling software	SCALEPACK
Phasing software	PHASER
Refinement software	CNS (1.1)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	50.000	2.450
High resolution limit [Å]	2.370	2.370
Number of reflections	46328
<I/σ(I)>	28.9	3.5
Completeness [%]	90.8	61.9
Redundancy	6.2

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	7.4	298	1.5 microliters of RebC (9 mg/mL in 150 mM NaCl, 10% glycerol, 25 mM HEPES pH 7.5) was incubated with 0.35 microliters of guanidine-HCl for 30 seconds, followed by addition of 1.5 microliters of precipitant solution (19% PEG-8000, 0.1 M HEPES pH 7.4), without mixing, at room temperature and sealed over a precipitant well solution. Immediately after set up, crystal trays were placed on a gel shaker and then, after 12 hours, transferred to a storage space in vibration-isolation. A crystal was then soaked in 19% PEG-8000, 0.1 M HEPES pH 7.4, and 5 mM chromopyrrolic acid for 1 week. The crystal was then soaked for 5 seconds in a cryogenic solution containing 19% PEG-8000, 0.1 M HEPES pH 7.4, 20% glycerol, and 5 mM chromopyrrolic acid and then flash-frozen in liquid nitrogen, VAPOR DIFFUSION, HANGING DROP, temperature 298K