4WED

Crystal structure of ABC transporter substrate-binding protein from Sinorhizobium meliloti

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 21-ID-D
Synchrotron site	APS
Beamline	21-ID-D
Temperature [K]	100
Detector technology	CCD
Collection date	2013-12-14
Detector	MARMOSAIC 300 mm CCD
Wavelength(s)	1.07819
Spacegroup name	P 31
Unit cell lengths	57.327, 57.327, 132.063
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	49.650 - 2.350
R-factor	0.1667
Rwork	0.164
R-free	0.22940
Structure solution method	SAD
RMSD bond length	0.013
RMSD bond angle	1.541
Data reduction software	HKL-3000
Data scaling software	HKL-3000
Phasing software	HKL-3000
Refinement software	REFMAC (5.8.0073)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	50.000	50.000	2.390
High resolution limit [Å]	2.350	6.370	2.350
Rmerge	0.057	0.038	0.777
Rmeas	0.065	0.043	0.913
Rpim	0.031	0.021	0.470
Total number of observations	86540
Number of reflections	19751
<I/σ(I)>	15.6
Completeness [%]	97.3	97.2	74.3
Redundancy	4.4	4.2	3.6
CC(1/2)		0.998	0.662

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	7.5	289	0.2 ul of 18 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-II condition # 18 (0.2 M Sodium Formate, 20% (w/v) PEG 3350 ) and equilibrated against 2.0 M NaCl solution in 96 Well 3 drop Crystallization Plate (Swissci). Before crystallization protein was incubated with 1/50 v/v of 2 mg/ml chymotrypsin solution at 289 K for 3 hours.