7R9X

Crystal structure of a dehydrating condensation domain, AmbE-CmodAA, involved in nonribosomal peptide synthesis

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 24-ID-E
Synchrotron site	APS
Beamline	24-ID-E
Temperature [K]	100
Detector technology	PIXEL
Collection date	2018-07-22
Detector	DECTRIS EIGER X 16M
Wavelength(s)	0.979
Spacegroup name	I 1 2 1
Unit cell lengths	89.940, 106.870, 108.909
Unit cell angles	90.00, 96.25, 90.00

Refinement procedure

Resolution	44.700 - 2.140
R-factor	0.1912
Rwork	0.190
R-free	0.23230
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	2jgp 6oyf
RMSD bond length	0.011
RMSD bond angle	1.123
Data reduction software	MOSFLM
Data scaling software	Aimless
Phasing software	PHENIX
Refinement software	PHENIX (1.15.2_3472)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	76.060	2.200
High resolution limit [Å]	2.140	2.140
Rmerge		0.257
Number of reflections	55789	55789
<I/σ(I)>	16.2	4.1
Completeness [%]	99.2
Redundancy	3.3
CC(1/2)	0.987	0.890

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP		277.15	AmbE_Cd crystallized at a concentration of 10mg/ml in conditions containing 100mM bis-Tris propane (pH 6.0), 20% PEG3350 and 0.2M sodium iodide. Cryoprotection was performed by dipping the crystal in a solution containing 20% MPD (hexylene glycol), 0.2M NaCl, 50mM Tris-Cl (pH7.5), 0.2M sodium iodide, 20% PEG 3350 and 100mM bis-Tris propane (pH 6.0) before being flash frozen in liquid nitrogen.