3HZS

S. aureus monofunctional glycosyltransferase (MtgA)in complex with moenomycin

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	ROTATING ANODE
Source details	RIGAKU FR-E DW
Temperature [K]	100
Detector technology	CCD
Collection date	2007-09-12
Detector	RIGAKU SATURN 944
Wavelength(s)	1.54
Spacegroup name	H 3 2
Unit cell lengths	114.422, 114.422, 128.844
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	23.120 - 2.100
R-factor	0.19183
Rwork	0.189
R-free	0.24379
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	Unpublished apo structure of MTGA
RMSD bond length	0.019
RMSD bond angle	1.800
Data reduction software	HKL-2000
Data scaling software	HKL-2000
Phasing software	AMoRE
Refinement software	REFMAC (5.4.0069)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	23.120	2.200
High resolution limit [Å]	2.100	2.100
Number of reflections	45531
<I/σ(I)>	13.6	3.2
Completeness [%]	99.9	100
Redundancy	3.9	3.8

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	4.6	295	Sa MtgA E100Q at 10 mg/ml was mixed with 1mM Moenomycin and 1mM MnCl2 and incubated on ice for ~3 hours. Precipitated material was removed by centrifugation at 16,000xg for 5 minutes. The Mmoenomycin complex was crystallized by hanging drop vapor diffusion, mixing the protein 1:1 with a reservoir solution containing 0.1M Na Acetate pH 4.6, 0.2M NaCl, 30% MPD at 22oC. Hexagonal plate crystals formed in 1 week., VAPOR DIFFUSION, SITTING DROP, temperature 295K