4UAM

1.8 Angstrom crystal structure of IMP-1 metallo-beta-lactamase with a mixed iron-zinc center in the active site

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	AUSTRALIAN SYNCHROTRON BEAMLINE MX1
Synchrotron site	Australian Synchrotron
Beamline	MX1
Detector technology	CCD
Collection date	2013-03-14
Detector	ADSC QUANTUM 210r
Wavelength(s)	1.283500
Spacegroup name	P 1
Unit cell lengths	49.990, 75.910, 82.360
Unit cell angles	83.45, 75.30, 74.01

Refinement procedure

Resolution	44.927 - 1.800
R-factor	0.1625
Rwork	0.162
R-free	0.18200
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	2doo
RMSD bond length	0.006
RMSD bond angle	0.962
Data reduction software	XDS
Data scaling software	XSCALE
Phasing software	PHASER
Refinement software	PHENIX (1.9_1692)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	44.930		1.850
High resolution limit [Å]	1.800	8.050	1.800
Rmerge	0.055	0.019	0.481
Rmeas		0.027	0.569
Number of reflections		1961	13902
<I/σ(I)>	18.08	42.02	2.9
Completeness [%]	94.8	83.7	92.26
Redundancy	3.9		3.9

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	6.5	297	drop contained 1ul protein (25-30mg/ml in 20mm HEPES, 50mm NaCl, ph7.5) with 1uL microseed reservoir (0.2M sodium acetate, 0.2M sodium citrate, 26% PEGMME 2K). Protein solution was heat-treated at 310K for 3-5 days prior to filtration and crystallisation screening. microseeding was essential for diffraction quality crystal growth. crystal grew over 2 weeks
1	VAPOR DIFFUSION, HANGING DROP	6.5	297	drop contained 1ul protein (25-30mg/ml in 20mm HEPES, 50mm NaCl, ph7.5) with 1uL microseed reservoir (0.2M sodium acetate, 0.2M sodium citrate, 26% PEGMME 2K). Protein solution was heat-treated at 310K for 3-5 days prior to filtration and crystallisation screening. microseeding was essential for diffraction quality crystal growth. crystal grew over 2 weeks