1R30

The Crystal Structure of Biotin Synthase, an S-Adenosylmethionine-Dependent Radical Enzyme

Experimental procedure

Experimental method	MAD
Source type	SYNCHROTRON
Source details	NSLS BEAMLINE X25
Synchrotron site	NSLS
Beamline	X25
Temperature [K]	100
Detector technology	CCD
Detector	ADSC QUANTUM 315
Wavelength(s)	1.30000, 1.73827, 1.74150
Spacegroup name	P 31 2 1
Unit cell lengths	155.690, 155.690, 90.880
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	44.450 - 3.400
R-factor	0.256
Rwork	0.256
R-free	0.30000
Structure solution method	MAD
RMSD bond length	0.009
RMSD bond angle	22.800 *
Data scaling software	XDS
Phasing software	SOLVE
Refinement software	CNS (1.0)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	100.000 *	3.510
High resolution limit [Å]	3.400	3.400
Rmerge	0.066 *	0.259 *
Number of reflections	17465
<I/σ(I)>	15.79	3.78
Completeness [%]	98.1 *	87.9 *
Redundancy	5.4 *	1.7

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	8 *	298	Tris, MgCl2, PEG 1000, Glycine, pH 6.5, VAPOR DIFFUSION, HANGING DROP, temperature 298.0K

Crystallization Reagents in Literatures

ID	crystal ID	solution	reagent name	concentration (unit)	details
1	1	drop	protein	20 (mg/ml)
2	1	drop	Tris-HCl	25 (mM)	pH8.0
3	1	reservoir	Tris-HCl	0.1 (M)
4	1	reservoir	glycine	0.1 (M)
5	1	reservoir		0.2 (M)
6	1	reservoir	PEG1000	20 (%(w/v))	pH6.5
7	1	reservoir		2.5 (M)