5BP7

Crystal structure of SAM-dependent methyltransferase from Geobacter sulfurreducens in complex with S-Adenosyl-L-homocysteine

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 21-ID-G
Synchrotron site	APS
Beamline	21-ID-G
Temperature [K]	100
Detector technology	CCD
Collection date	2015-04-16
Detector	MARMOSAIC 300 mm CCD
Wavelength(s)	0.97856
Spacegroup name	P 21 21 2
Unit cell lengths	89.346, 110.189, 76.681
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	50.000 - 2.100
R-factor	0.1873
Rwork	0.185
R-free	0.22290
Structure solution method	SAD
RMSD bond length	0.010
RMSD bond angle	1.458
Data reduction software	HKL-3000
Data scaling software	HKL-3000
Phasing software	HKL-3000
Refinement software	REFMAC (5.8.0107)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	50.000	50.000	2.140
High resolution limit [Å]	2.100	5.700	2.100
Rmerge	0.123	0.064	0.995
Rmeas	0.130	0.070
Rpim	0.054	0.029	0.444
Total number of observations	276539
Number of reflections	45102
<I/σ(I)>	5.1		1.6
Completeness [%]	99.9	99.6	100
Redundancy	6.1	5.5	6
CC(1/2)		0.997	0.756

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	7.5	289	0.2 ul of 15 mg/ml protein in 20 mM HEPES pH 7.5, 150 mM NaCl, 10% Glycerol, 0.1% Sodium Azide and 0.5 mM TCEP were mixed with 0.2 ul of the MCSG Suite 1 condition #9 (0.1M HEPES, 25%w/v PEG 3350, 0.2M Magnesium Chloride, 6-Hydrate pH=7.5) and equilibrated against 1.5 M NaCl solution in 96 Well 3 drop Crystallization Plate (Swissci). Before crystallization protein was incubated with 5 mM S-adenosylmethionine chloride and 1/50 v/v of 1 mg/ml TEV solution at 289 K for 3 hours