4KS6

Crystal structure of the catalytic domain of botulinum neurotoxin BoNT/A C134S mutant with covalent inhibitor that modifies Cys-165 causing disorder in 166-174 stretch

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ESRF BEAMLINE ID23-1
Synchrotron site	ESRF
Beamline	ID23-1
Temperature [K]	100
Detector technology	CCD
Collection date	2011-06-13
Detector	ADSC QUANTUM 315r
Wavelength(s)	0.97240
Spacegroup name	P 43 21 2
Unit cell lengths	65.634, 65.634, 201.725
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	62.410 - 1.930
R-factor	0.19561
Rwork	0.193
R-free	0.25199
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	4elc
RMSD bond length	0.020
RMSD bond angle	1.514
Data reduction software	XDS
Data scaling software	XDS
Phasing software	REFMAC
Refinement software	REFMAC

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	50.000	50.000	1.900
High resolution limit [Å]	1.790	5.340	1.790
Rmerge	0.377	0.060	4.886
Number of reflections	42323
<I/σ(I)>	9.79	37.96	0.86
Completeness [%]	99.2	99.6	94.9
Redundancy	15.4	14.02	15.42

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	6	293	protein complex - 6 microL LCA C134S at 145 microM + 1.4 microL KG23 at 1.25 milli-M, Reservoir - 17.5% PEG 600, 0.065 M HEPES pH 7.5. Cryoprotectant - CryoProtX CM1, 10% PEG 20K, 100 milliM CHC (citric acid, HEPES, CHES - 40% pH 4 / 60% pH 10), VAPOR DIFFUSION, SITTING DROP, temperature 293K